BioResearch Bio Research Cancer Research Tools for Biological Relevance Os nos ad tat. Igna faccum eroses iriusti onulla feuisisit adio con veliquamcore magna alis nonulla non er irilla am, quiscil dolobor eu faccum quisisim quis nullaor rem iril er summy nulpute tinos ipisci te facilit nulla facilis molore. Cancer Research and Drug Discovery Tools BioResearch Cancer Research and Drug Discovery Tools Avoid Research Roadblocks and Take a Direct Route to Results “I need to understand the origin of cancer and what role cancer stem cells play in cancer formation” SOLUTION: Cancer stem cell hypothesis and cancer formation can be simulated optimally in well-characterized primary cells. Page 5 Primary Cells and Media “Efficient transfection of various cancer cell lines or primary cells with plasmids or siRNA requires a lot of optimization effort.” SOLUTION: Pick from more than 100 ready-to-use Nucleofection™ Protocols suited for high efficiency with any nucleic acid substrate. Page 8 Transfection 2 “I want to use primary cells for high throughput drug screening, but can't get large quantities of homogenous primary cells.” SOLUTION: Lonza offers conditionally immortalized cells from a wide range of human tissues that offer primary-like characteristics. Page 13 Drug Discovery Tools “I can’t find reliable assay tools to study drug effects on protein expression, cell proliferation, or cytotoxicity.” SOLUTION: Choose from our variety of assay tools, including ready-to-use protein gels and bioassay kits to measure cell proliferation and cytotoxicity. Page 14 Cell Analysis 3 BioResearch Cancer Research and Drug Discovery Tools Your Cancer Research Toolkit relevant results, from high-quality primary cells, through efficient transfection technology, to a wide range of moleculary biology and cell culture tools. Streamline your workflow by choosing from convenient, innovative research tools that have been designed and tested to work together. Lonza gives you what you need for biologically Primary Cells and Media Clonetics™ Human Cells with optimized growth media: Blood & Lymph Cells – Blood & lymphatic HMVECs – Hematopoietic stem cells – Mesenchymal stem cells Bone Cells –Osteoblasts – Osteoclast precursors –Fresh bone marrow and progenitor cells Breast Cells – Mammary epithelial Pancreatic Cells – Pancreatic Islets – Acinar tissue Pulmonary Cells –Bronchial epithelial & smooth muscle – Lung Fibroblasts and HMVECs –Pulmonary artery endothelial and smooth muscle – Small airway epithelial cells Reproductive and Renal Cells –Prostate epithelial, smooth muscle and stromal cells –Uterine and Bladder endothelial and smooth muscle cells – Sertoli cells Skin Cells – Adult and neonatal HMVECs –Fibroblasts –Keratinocytes –Melanocytes –Pre-adipocytes and Adipose-derived stem cells Transfection Drug Discovery Tools Cell Analysis Nucleofector™ Technology with optimized protocols for cancer-related primary cells, including: – Blood cells – Epithelial cells – Endothelial cells – Neuronal cells –Fibroblasts – Adult or embryonic stem cells Conditionally Immortalized Human Cell Lines: –Brain Microvascular Endothelial Cells –Preadipocytes – Skeletal Muscle Cells – Dermal Fibroblasts –Coronary Artery Smooth Muscle Cells (CASMC) –Blood (BEC) and Lymphatic (LEC) Microvascular Endothelial Cells – Adult Dermal Keratinocytes Cell Proliferation and Cytotoxicity BioAssay Kits: –ViaLight™ Kit for measuring cell proliferation and cytotoxicity – ToxiLight™ Kit for non-destructive measurement of cytotoxicity Also optimized for more than 80 cancer-related cell lines, including: – Blood cancer (e.g. Jurkat) – Breast cancer (e.g. MCF-7) – Lung cancer (e.g. A549) – Neurological cancer (e.g. C6) – Prostate cancer (e.g. DU145) Biowhittaker™ Liquid and Powder Media for therapeutic and research needs Cell Function Assays: –Bone mineralization and resorption assays –Adipogenesis and lipolysis measurement assays Mycoplasma Detection: –MycoAlert™ Assay for accurate, reliable and universal mycoplasma detection DNA, RNA, and Protein Expression Analysis: –PAGEr™ EX Gels and ProSieve™ EX Reagents for protein analysis and Western blotting –FlashGel™ System for PCR analysis and RNA integrity analysis 4 Primary Cells and Media We offer a wide variety of cells and optimized media kits for cancer research. Cells and media are tested together and guaranteed to perform. “I need to understand the origin of cancer and what role cancer stem cells play in cancer formation” Primary cells are an essential component in research because they are derived directly from human and animal tissue representing a living, biologically relevant cell model. Primary cells have low modification/ mutation rate and require minimal pre-characterization prior to use. We offer over 100+ primary cell types and optimized media kits for each. Cells and media are tested together and guaranteed to perform. SOLUTION: Cancer stem cell hypothesis and cancer formation can only be simulated optimally in well-characterized primary cells. Benefits of Primary Cells –– Better alternatives to cell lines Research shows, some cell lines may not be the cell type they have historically been reported. In a study of over 500 leukemialymphoma cell lines, 15% of the cell lines were misidentified.1 In a separate study, 18-36% of cell lines are found to be misidentified or cross-contaminated.2 For this reason, government institutions such as NIH mandates authentication of cultured cell lines as critical for grant applications to be considered of the highest quality.3 Primary cells could be used as both normal controls to extensively characterize and validate your cell lines or they may solely act as superior substitutes to cell lines. –– Biologically relevant for biomarker discovery Cell lines are prone to genotypic and phenotypic drift after continual passaging and many journals and government funding programs mandate authentication of cell lines prior to use in experiments. In addition, between 3-10 cell lines are required from same origin to build a comprehensive biomarker profile. This can drive up experimental costs significantly. Clonetics™ Primary Cells are extensively validated and make biomarker discovery more comprehensive as these retain cell surface markers and other genes relevant to actual human tissue. Eliminate costly analyses of inconsistent data acquired from non-characterized cell lines or low-quality primary cells. –– Ideal for discovery of anti-cancer drugs Clonetics™ Primary Cells have successfully been published in drug screening assays. They are ideal solution for testing new anticancer drugs as they are representative of actual human system. The preclinical relevance of primary cells is becoming increasingly significant in cancer therapeutics. Models developed in primary cell culture can easily be replicated in actual animal studies. In addition, drug resistance assays for chemotherapy have successfully been refined by researchers using primary cells alone. Reduce the cost and number of animals required for in vivo studies by refining your experiments with primary cells first (figure1). –– Understand how cancer forms Research shows primary cells can successfully be transformed into cancer cells through genetic mutations and expression of oncogenes. Replicate a cancer model in vitro using primary cells as prototypes of in vivo environment. –– Study Cancer Stem Cells Cancer stem cells (CSCs) are stem-cell like components that originate from normal tissue stem cells. CSCs share several properties with normal stem cells, such as their capacity for self-renewal and their ability to differentiate. CSCs, currently, can only be studied in primary cells or normal tissues as both contain sub-populations of stem-cell markers. Research also shows primary cells can be reprogrammed into iPSCs and this is a vast area of study in cancer research currently. 1) Drexler HG, Dirks WG, Matsuo Y and MacLeod RAF (2003) Leukemia 17, 416–426 2) Hughes P et al. (2007) Biotechniques 43:575-586 3)http://grants.nih.gov/grants/guide/notice-files/NOT-OD-08-017.html 5 BioResearch Cancer Research and Drug Discovery Tools % maximal response Primary Cells and Cell Lines Show Variability in Drug Dose Response 110 100 90 80 70 60 50 40 30 20 10 0 Some Pathways Can Only Be Studied in Primary Cells Isolated CTL 1 HEK 293 EC 50 2 primary HRE CTL docked to target cell 3 [Camptothecin] nM primary HRCE HEK 293 HRE HRCE 68.10 537.5 557.4 Figure 2. Some pathways can only be studied in primary cells as there is no cell line to represent that pathway. Cytolytic T lymphocytes (CTLs) recognize foreign cells and lyse them via the “immunological synapse”. At the immunological synapse CTLs secrete cytolytic granules containing Perforin. hMUNC13-4 is involved in an activation step preceding vesicle membrane fusion in cytolytic granule secretion. Nucleofection™ of a plasmid expressing hMUNC13-4/RFP fusion shows only partial co-localization of hMUNC13-4 (red, H) and Perforin (green, I) in isolated CTLs (overlay J; blue: nuclei stained by DAPI). Upon target cell contact (M) hMUNC13-4 polarizes and extensively co-localizes with Perforincontaining cytolytic granules at the immunological synapse (L). Reprinted from: Feldmann J et al. (2003) Cell 115(4):461-73; with permission of Elsevier. Figure 1. Primary cells and cell lines show variability in drug dose so data acquired through cell lines cannot easily be replicated in an in vivo model. The three cell types were dosed with Camptothecin for 48h and their cytotoxic response assessed using the ViaLight™ Assay.The ViaLight™ Assay measures the number of viable cells left after drug challenge, measuring the ATP donated to the assay reaction by viable cells. In this assay the data points are the mean of 40 replicates (8 replicates per plate; 5 plates). We have a marked difference in EC50 dose response almost 10 fold. The HEK 293 cell line required only 68nM Camptothecin to kill 50% of the cells. In the case of the primary cells (HRE = human renal epithelial cells, HRCE = human renal cortical epithelial cells) this dose was over 500nM a striking different response. 6 Clonetics™ Primary Cells Poietics™ Adult Stem Cells and Media Why use Clonetics™ Primary Cells? − Product quality – Superior to cell lines with minimum specs gua rantee. All Clonetics™ Cells are ISO-certified with extensive quality control and batch-to-batch consistency −Product availability – Readily available 100+ cell types (human and animal) with donor variety and steady supply schedule − Reliability – Market leader in primary cells offering; 40+ years of experience and 10× more publications than other providers − Cost-effective – No pre-characterization needed; No repeat analyses; Refine experiments and reduce animal work − Easy to culture – Specialized support team to assist, optimized protocols guaranteed to work − Convenience – Cryopreserved ampoule to store and use as needed or proliferating in flasks/plates to save time All of Lonza’s stem cells are guaranteed to be of high purity and possess full differentiation capacity. –– Studies have shown that mesenchymal stem cells can contribute to cancer progression in a number of different cancers, primarily cancers affecting bone marrow, blood and lymph tissues. Lonza offers cryopreserved human and rat Mesenchymal Stem Cells, as well as optimized Mesenchymal Stem Cell Growth and Differentiation media. –– Lonza has a variety of human hematopoietic stem cells from cord blood and Bone marrow, along with specialized growth medium and EasyDiff™ kits designed to differentiate CD34+ progenitors into Erythroid, Megakaryocyte and Myeloid Cell. –– Lonza also offers Osteoclast Precursors, Preadipocytes and Adipose-Derived stem cells, along with specialized media for each, that are perfect for studying various bone and lipoma tumors. –– Lonza runs its own bone marrow donor program to collect normal bone marrow aspirates from healthy donors for use in research. This program provides a convenient, easy to access source of bone marrow derived cells. Case Study: Clonetics™ Human Prostate Epithelial Cells (PrECs) and Media in Cancer Stem Cells Applications Isotype A Counts The Role of CD133 in Normal Human Prostate Stem Cells and Malignant Cancer-Initiating Cells. D. J. Vander Griend et.al. (2008) Cancer Res 68:9703-9711 ABCG2 CD133 200 160 120 80 40 101102103104 IgG-PE B 104 101102103104 CD133-PE –– Prostate cancer arises within the epithelial compartment and generally believed to begin within the stem-cells –– In the adult human prostate, CD133/ABCG2 expression is indicative of small population of putative stem cells that suppress differentiation −Clonetics™ PrECs (Cat. No. CC-2555) were utilized alongside inhouse isolated PrEC’s to assess stem-cell like properties. Both cell types were cultured in Clonetics™ PrEGM Growth Media BulletKit (Cat. No. CC-3166). Both exhibit high expression of stem-cell markers CD133/ABCG2 (figure 3) 103 104 103 104 104 105 105 101102103104 IgG-FITC 101102103104 101102103104 ABCG2-FITC Figure 3. Expression of stem cell markers in PrEC cultures. A: ABCG2-FITC and CD133-PE (AC141) expression in PrECs. B: Dual-label flow cytometry showing that CD133+ cells are also ABCG2+ Reprinted from Cancer Res, 2008, 68:9703-9711, DJ Vander Griend et al, The Role of CD133 in Normal Human Prostate Stem Cells and Malignant Cancer-Initiating Cells, with permission from AACR. 7 BioResearch Cancer Research and Drug Discovery Tools Transfection of Cancer-related Primary Cells and Cell Lines As the leading non-viral technology for transfection of difficult-to-transfect cell lines and primary cells, the Nucleofector™ Technology is an important tool to help understand the up-regulation and down-regulation of genes in the progression from normal to cancerous cells and identifying potential drug targets. “Efficient transfection of various cancer cell lines or primary cells with plasmids or siRNA requires a lot of optimization effort.” SOLUTION: Pick from more than 100 ready-to-use Nucleofection™ Protocols suited for high efficiency with any nucleic acid substrate. Benefit From Applications –– Study the role of genes in the progression from normal to cancerous cells –– Identification of potential drug targets –– Confirmation of findings based on a cell line in further cell lines or even corresponding primary cells –– Easy switch from overexpressing genes (via DNA, RNA or peptides) to down-regulation (by siRNA or shRNA) Blood Cancer Breast cancer Colon Cancer Liver Cancer Neurological Cancer Prostate Cancer MG63 HeLa Human PrEC PC3 LnCAP DU 145 U-87MG Neuron, cortical, rat SH-SY5Y Neuro-2a [N2a] GH3 IMR32 C6 NHBE T84 Lung Cancer Figure 4. Examples of cancer cell lines and primary cells successfully transfected by Nucleofection™. Data shown represent average results from various data sets. 8 HMVEC-L A549 Hep G2 Hepatocyte, human HT29 HCT 116 HMEC T-47D MCF7 T cell, human (unstim.) U-937 B cell, human RAW 264.7 Raji Ramos K-562 Jurkat 100 90 80 70 60 50 40 30 20 10 0 HL-60 Average Transfection Efficiency (%) –– Non-viral transfection of primary cells and difficult-to-transfect cancer cell lines –– Up to 90% transfection efficiency and high cell viability –– High flexibility in cell types and substrates –– Different platforms for low throughput and high throughput, for e.g. siRNA screenings. –– More than 100 ready-to-use Nucleofection™ Protocols for a huge spectrum of primary cells and cancer cell lines, including blood, breast and prostate cancer –– More than 800 cancer-related publications reducing the effort to optimize methodologies –– Option of adherent Nucleofection™ adding more convenience and circumventing trypsinization Other Choose the Nucleofection™ Platform That Suits Your Research Needs Advanced Platform 96-well Add-on High-throughput Platform 4D-Nucleofector™ System 96-well Shuttle™ System 384-well Nucleofector™ System Nucleofector™ 2b Device Throughput (samples per run) Low to medium (1 – 16) Low to high (1 – 96) High (384) Low (1) Reaction volume 20 µl + 100 µl 20 µl 20 µl 100 µl Electrode material Conductive polymer Conductive polymer Conductive polymer Aluminum Low cell numbers (20 µl) 5 × 10 to 1 × 10 5 × 10 to 1 × 10 5 × 10 to 1 × 10 – High cell numbers (100 µl) 2 × 105 to 2 × 106 – – 2 × 105 to 2 × 106 DNA Vector amount/sample 0.2 – 1 μg (20 µl) 1 – 5 μg (100 µl) 0.2 – 1 μg 0.2 – 1 μg 1 – 5 μg siRNA amount/sample (concentration 2 nM – 2 µM) 0.04 – 40 pmol (20 µl) 0.2 – 200 pmol (100 µl) 0.04 – 40 pmol 0.04 – 40 pmol 0.2 – 200 pmol Adherent Nucleofection™ n – – – Compatibility with 96-well Shuttle™ System n – – – Device Basic Device Unit 4 5 4 9 5 4 5 BioResearch Cancer Research and Drug Discovery Tools Overview of Successfully Transfected Cell Types The table below shows a selection of cell types for which Nucleofection™ Protocols or data exist. Further cell types can be found in our cell database. www.lonza.com/cell-database Efficiency* Viability* Kit for 4D-Nucleofector™ and 96-well Shuttle™ Systems Kit for Nucleofector™ II/2b Device B cell, human 28–36% 84–92% P3 Human B Cell Dendritic cell, human 93–99% 12–75% Contact Scientific Support Human Dendritic Cell Macrophage, human 42–59% 87–88% P3 Human Macrophage Monocyte, human 60-64% 62–81% P3 Human Monocyte T cell, human stim. 41–70% 83–90% P3 Human T Cell T cell, human unstim. 43–98% 51–92% P3 Human T Cell AML 20–70% 45–87% Contact Scientific Support Contact Scientific Support B–CLL 30–60% 75–90% Contact Scientific Support Contact Scientific Support CML 42% 80% Contact Scientific Support Contact Scientific Support EL4 (Mouse T cell lymphoma) 65% 76% SF Cell Line L HL-60 (Human acute myeloid leukemia) 90% 50–65% SF Cell Line V Jurkat (Human T cell leukemia) 88% 90% SE Cell Line V K-562 (Human CML-derived B cell line) 80–90% 88% SF Cell Line V KG-1 (Human AML cell line) 70% 84% SE/SF/SG Cell Line R Raji (Human Burkitt’s lymphoma) 84% 67–81% SG Cell Line V Ramos (Human Burkitt’s lymphoma) 27–51% 70–77% SG Cell Line V RAW 264.7 (Mouse macrophage leukemia) 60–65% 74–86% SF Cell Line V THP-1 (Human acute monocytic leukemia) 47–68% 40–81% SG Cell Line V U-937 (Human histiocytic lymphoma) 32–67% 62–89% SG Cell Line C 50–73% 66–98% P3 HMEC MCF7 (Human mammary gland adenocarcinoma) 72–78% 60% SE Cell Line V MDA-MB-231 (Human breast adenocarcinoma) 73–89% 77% SE Cell Line V MDA-MB-453 (Human breast adenocarcinoma) 54% 90% SE/SF/SG Cell Line C MDA-MB-468 (Human breast adenocarcinoma) 60% 81% SE/SF/SG Cell Line V SK-BR-3 (Human breast adenocarcinoma) 50% 94% SE/SF/SG Cell Line C T-47D (Human mammary gland ductal carcinoma) 51-80% 94% SE Cell Line V Caco-2 (Human colorectal adenocarcinoma) 59–82% 70–80% SE Cell Line T HCT 116 (Human colorectal adenocarcinoma) 70–80% 65–76% SE Cell Line V HT-29 (Human colorectal adenocarcinoma) 16–67% 57–94% SF Cell Line R SW480 (Human colorectal adenocarcinoma) 60% 86% SE/SF/SG Cell Line V Blood Cancer – Primary Cells Blood Cancer – Cell Lines Breast Cancer – Primary Cells Epithelial, mammary (HMEC), human Breast Cancer – Cell Lines Colon Cancer – Cell Lines 10 Efficiency* Viability* Kit for 4D-Nucleofector™ and 96-well Shuttle™ Systems Kit for Nucleofector™ II/2b Device 60–84% P3 Human Chondrocyte Connective Tissue/Bone Cancer – Primary Cells Chondrocyte, human 65–75% Connective Tissue/Bone Cancer – Cell Lines MG-63 (Human osteosarcoma) 62–73% 60–90% SE Cell Line C Saos-2 (Human osteosarcoma) 82% 79% SE/SF/SG Cell Line V U-2 OS (Human osteosarcoma) 98% 88% SE/SF/SG Cell Line V Epithelial, kidney, human 40% 65% P3/P1 – Epithelial, renal proximal tubule cells (RPTEC), human 70% 90% P3/P1 – 54% 59–69% P3 – Hep G2 (Human hepatocellular carcinoma) 41–95% 86–94% SF Cell Line V Hep1B (Mouse hepatocarcinoma) 50–56% SE/SF/SG Cell Line R HuH7 (Human hepatoma) 60–90% 95% SE/SF/SG Cell Line T Endothelial, microvascular, lung (HMVEC–L), human 52–79% 48–52% P5 HMVEC-L Epithelial, bronchial (NHBE), human 50–65% 50–53% P3 Basic Epithelial Cell A549 (Human lung carcinoma) 72–81% 62–81% SF Cell Line T Calu-3 (Human lung adenocarcinoma) 40–81% 54–90% SE Cell Line V Calu-6 (Human lung carcinoma) 73% 50–87% SE/SF/SG Cell Line L IMR-90 (Human lung embryonic fi broblasts) 51–65% 70% SE Cell Line R NCI-H1299 [H1299] (Human lung cancer) 99% 75% SE/SF/SG Cell Line C T84 (Human lung metastase from colorectal carcinoma) 53–88% 50–83% SF Cell Line T Kidney Cancer – Primary Cells Liver Cancer – Primary Cells Hepatocytes, human Liver Cancer – Cell Lines Lung Cancer – Primary Cells Lung Cancer – Cell Lines Neurological Cancer – Primary Cells Astrocyte (NHA), human 83–85% P3 Basic Glia Cell Astrocyte, mouse 60% 60–70% P3 Basic Glia Cell Neuron, cerebellar granule, mouse 30–51% 80% P3 Mouse Neuron Neuron, cerebellar granule, rat 50–60% 50–70% P3 Rat Neuron Oligodendrocyte, rat 44% 60% P3 Basic Glia Cell Neuroblastoma 20–90% P3 contact Scientific Support 11 Continued on next page BioResearch Cancer Research and Drug Discovery Tools Efficiency* Viability* Kit for 4D-Nucleofector™ and 96-well Shuttle™ Systems Kit for Nucleofector™ II/2b Device C6 (Rat glioma) 92–94% 55–80% SF Cell Line V GH3 (Rat pituitary tumor) 60–80% 60–84% SE Cell Line L IMR-32 (Human neuroblastoma) 74–86% 45–63% SF Cell Line L Neuro-2a [N2a] (Mouse neuroblastoma) 46–90% 81–97% SF Cell Line V SH-SY5Y (Human neuroblastoma) 63–82% 40–80% SF Cell Line V SKNAS (Human neuroblastoma) 41–57% 55–82% SE/SF/SG Cell Line T SK-N-MC (Human neuroblastoma) 50–95% 30–60% SF Cell Line V SK-N-SH (Human neuroblastoma) 85% 73% SE/SF/SG Cell Line V CHO-K1 (Hamster ovary) 76–94% 53–97% SF Cell Line T HeLa (Human cervical adenocarcinoma) 60–85% 80–89% SE Cell Line R OVCAR3 (Human ovarian adenocarcinoma) 70–80% 70–95% SE/SF/SG Cell Line T SK-OV-3 (Human ovarian adenocarcinoma) 89% 53% SE/SF/SG Cell Line V 43–67% 48–64% P1 Basic Epithelial Cell DU 145 (Human prostate carcinoma) 47–100% 70–92% SE Cell Line L LNCaP (Human prostate carcinoma) 70–82% 45–80% SF Cell Line R PC-3 (Human prostate adenocarcinoma) 83–88% 59–66% SF Cell Line V Fibroblast, dermal (NHDF), human – adult 42–96% 74–100% P2 Human Fibroblast Fibroblast, dermal (NHDF), human – neo 90–98% 85–91% P2 Human Fibroblast Keratinocyte, adult (NHEK–Ad), human 51% 40–60% Primary Cell Optimization Human Keratinocyte Keratinocyte, neonatal, pooled (NHEK–neo), human 39–78% 50–79% P3 Human Keratinocyte Melanocyte, neonatal (NHEM–neo), human 70% 55–60% Primary Cell Optimization Human Melanocyte A2058 (Human skin melanoma) 81% 94% SE/SF/SG Cell Line C A-375 (Human skin melanoma) 72% 97% SE/SF/SG Cell Line V B16-F0 (Mouse skin melanoma) 84% 91% SE/SF/SG Cell Line R CD34+ cell, bone marrow, human 82% 62–70% P3 Human CD34+ Cell Embryonic stem (ES) cell, human 20–78% 50–98% P3 Human Stem Cell Mesench. stem (MSC), human 55–88% 50–86% P1 Human MSC Neural stem cell (NSC), human 90% P3 or P4 Basic Neuron Neural stem cell (NSC), mouse 60–82% P4 Mouse NSC Neural stem cell (NSC), rat 42–46% P3 Rat NSC Neurological Cancer – Cell Lines Ovarian/Cervical Cancer – Cell Lines Prostate Cancer – Primary Cells Epithelial, prostate (PrEC), human Prostate Cancer – Cell Lines Skin Cancer – Primary Cells Skin Cancer – Cell Lines Stem Cells 80% * Approximate ranges extrapolated from larger result collections, including Lonza and customer data 12 Drug Discovery Tools Clonetics™ Immortalized Cells (CCICs) – Models for Screening Anti-Cancer Drugs “I want to use primary cells for high throughput drug screening, but can't get large quantities of homogenous primary cells.” CCICs present a solution to drug screening with biologically relevant cells. These cells maintain most of the valuable properties of a primary cell but possess practically unlimited growth potential, thus representing an ideal model for cell based high throughput screening. Screen cells for biomarkers and cancer drug targets efficiently with large volumes of homogenous primary-like cell populations. SOLUTION: Lonza offers conditionally immortalized cells from a wide range of human tissues that offer primary-like characteristics. (% over basal) We offer following cell lines in our CCIC portfolio –– Human Brain Microvascular Endothelial Cells –– Human Preadipocytes –– Human Skeletal Muscle Cells –– Human Dermal Fibroblasts –– Human Coronary Artery Smooth Muscle cells (CASMC) –– Human Blood (BEC) and Lymphatic (LEC) Microvascular Endothelial Cells –– Human Adult Dermal Keratinocytes 400 A Passage 6 300 200 100 0 Classical & Specialty Media (% over basal) The Biowhittaker™ brand provides over 16 high quality media products that fit a variety of cancer applications. Our highly referenced media portfolio contains a variety of liquid and powder formulations that have been supporting therapeutic and research needs for over 20 years. The BioWhittaker™ brand includes classical media, reagent, or regulatory friendly specialty media and is manufactured in accordance with cGMP regulations. 400 Basal -9 -8 -7 -6 [Insulin] M B Passage 21 300 200 100 0 www.lonza.com/culturemedia Basal -9 -8 -7 -6 [Insulin] M Figure 5. Conditionally immortalized skeletal muscle cells maintain the ability for insulin dependent glycogen synthesis after extended passaging. XM15B1 myoblasts were serum starved for 18 h in Ham’s F10 (Lonza) medium at 37°C and exposed to insulin for 2 h in serum free medium containing radiolabeled glucose. The synthesized glycogen was quantified using a filter based radiometric assay. Cells show a dose dependent increase in glycogen synthesis in response to insulin at early (A, passage 6) and late (B, passage 21) passages. 13 BioResearch Cancer Research and Drug Discovery Tools Cell Analysis Tools for Cancer Research Utilize Lonza’s research tools for every step of your workflow. Lonza provides everything you need for biologically relevant results – from culture media, sample preparation, all the way through protein confirmation. "I can’t find reliable assay tools to study drug effects on, protein expression, cell proliferation, or cytotoxicity.” ViaLight™ Plus Kit SOLUTION: Choose from our variety of assay tools, including ready-to-use protein gels, and bioassay kits to measure cell proliferation and cytotoxicity. Measure cell proliferation and cytotoxicity by using stable bioluminescence to detect adenosine triphosphate (ATP): –– Process and analyze a 96-well plate in less than 15 minutes –– Detect as few as ten cells; allowing for lower seeding densities and more assays –– Easily perform scalable automation with our simple, no-shake protocol –– Add two reagents directly to your culture and read luminescence –– Expand your dynamic range to five decades, in adherent or suspension cultures –– Run this test on a variety of luminometers or scintillation counters –– Avoid radioactive or toxic materials AdipoLyze™ Lipolysis Detection Assay The AdipoLyze™ Lipolysis Detection Assay is a fast, sensitive, fluorescent in vitro assay used for detecting small quantities of glycerol in cells undergoing lipolysis. –– Quantify in vitro lipolysis of adipogenic cell lines and both subcutaneous and visceral primary preadipocytes –– Accurately detect very low levels of glycerol (down to 0.44 μM or 0.04 μg/ml) www.lonza.com/vialight ToxiLight™ Kit AdipoRed™ Assay Reagent Check a compound’s cytotoxic effects non-destructively by measuring adenylate kinase (AK) leakage from damaged cells: –– Generate results from as few as 10 cells –– Eliminate the need to lyse by monitoring cytotoxicity from supernatant –– Simply add a single reagent directly to cells, or to a supernatant aliquot –– Process and analyze a 96-well plate in less than 10 minutes –– Freeze supernatants and return to your work later without losing AK activity The AdipoRed™ Assay Reagent is designed for assessing the effect of compounds on the differentiation of preadipocytes or on lipid utilization in mature adipocytes. This objective, high-throughput, homogeneous fluorescence-based assay quantifies the accumulation of intracellular triglycerides and provides significant advantages to drug discovery efforts. − More sensitive than other methods such as the Oil Red O assay − Faster and easier than Northern and Western blots www.lonza.com/toxilight 14 OsteoAssay™ Human Bone Plate OsteoImage™ Mineralization Assay The OsteoAssay™ Plate provides a thin layer of adherent real human bone for the culture of primary human or non-human osteoclasts, osteoclast precursors, or immortalized cell lines. Cells can be stained with standard cytochemical or immunofluorescent techniques. Assays of bone resorption and osteoclast precursor differentiation can be performed easily by sampling the cell culture supernatant. The OsteoImage™ Mineralization Assay is a rapid, fluorescent in vitro assay for assessing bone cell mineralization. The OsteoImage™ Assay can quantitate in vitro mineralization by osteogenic stem cells, primary osteoblasts, and osteoblast-like cell lines. It is based on specific binding of the fluorescent OsteoImage™ Staining Reagent to the hydroxyapatite portion of bone-like nodules deposited by cells. Unlike typical histochemical methods such as von Kossa and Alizarin red, neither of which is hydroxyapatite specific, the OsteoImage™ Assay eliminates multiple steps or tedious extraction steps. –– –– –– –– –– Eliminates the need for dentine or animal bone slices Simply seed cells onto the surface of the OsteoAssay™ Plate Use with a variety of cell types and cell-based assays Contains real human bone for more biologically relevant results –– Delivers qualitative, visual fluorescent microscopy or quantitative plate reader results –– Can be used with primary osteoblasts, osteoblast stem cells, and osteoblast cell lines –– Measures hydroxyapatite, similar to real bone –– Completed in less than 90 minutes, without tedious extractions –– Sensitive enough to detect time-dependent increases in mineralization in differentiating cells –– Scalable for use in 6-well up to 96-well plates OsteoLyse™ Assay Kit The OsteoLyse™ Assay Kit provides an easy to use protocol for quantitatively measuring in vitro osteoclast-mediated bone resorption in a high-throughput format. Osteoclasts can be seeded onto the OsteoLyse™ plate using traditional cell culture protocols. The assay directly measures the release of Europium-labeled collagen fragments into the osteoclast cell culture supernatant via time resolved fluorescence, indicating their resorption activity levels. MycoAlert™ Mycoplasma Detection Assay The MycoAlert™ Assay Kit is a selective biochemical test that exploits the activity of specific mycoplasmal enzymes which are found in all 6 of the main mycoplasma cell culture contaminants and the vast majority of 180 species of mycoplasma. –– No need to separately purchase bone matrices as human collagen is bound to wells –– Just seed cells onto the OsteoLyse™ Plate surface –– Measure resorptive activity by simply sampling the cell culture supernatant and counting via time-resolved fluorescence –– Results in <20 minutes by a simple 2-step assay –– Bioluminescence-based technology – no DNA extraction necessary –– NEW! MycoAlert™ PLUS Kit for use with less sensitive plate luminometers or multifunctional readers www.lonza.com/mycoalert 15 BioResearch Cancer Research and Drug Discovery Tools Custom Research Solutions Molecular Biology Products Nucleic Acid Electrophoresis: – FlashGel System If you’re looking for customized research solutions or expert support services, just ask your local Lonza representative. They can put you in touch with experts that have decades of experience in cell isolation, transfection, and testing services. Protein Electrophoresis: – PAGEr™ EX Gels – ProSieve™ EX Reagents Custom Cell and Screening Services Lonza’s molecular biology products offer basic aspects of cancer research. Speed up your results from PCR analysis, RNA integrity analysis, to fragment isolation for cloning. Lonza’s FlashGel™ System can get you there in minutes. For protein analysis and western blotting Lonza’s PAGEr™ EX Gels and ProSieve™ EX Reagents help speed your research from hours to minutes. You can order virtually any human or animal cell through our vast tissue network. Lonza’s isolation experts can provide you with: –– A broad range of cells, packaged to your specifications –– Customized treatments, such as cell pelleting, personalized media, and specific treatment prior to cryopreservation Scientific Support For advice or information on any Lonza product, just contact our scientific support team. They offer: –– More than 50 years of combined troubleshooting experience –– Educational webinars –– Real disease research application data Y ou can find contact details for your regional support team on the back of this brochure. 16 Ordering Information Cell Type Description Cat# (cells) Recommended G rowth Media Media Bulletkit Cat # HMVEC-dBIAd Adult dermal blood microvascular endothelial CC-2811 EGM-2MV CC-3202 HMVEC-dBlNeo Neonatal dermal blood microvascular endothelial CC-2813 EGM-2MV CC-3202 HMVEC-dLyAd Adult dermal lymphatic microvascular endothelial CC-2810 EGM-2MV CC-3202 HMVEC-dLyNeo Neonatal dermal lymphatic microvascular endothelial CC-2812 EGM-2MV CC-3202 HMVEC-L Lung Microvascular Endothelial CC-2527 EGM-2MV CC-3202 HMVEC-Lly Lung lymphatic microvascular endothelial CC-2814 EGM-2MV CC-3202 CD133+ Hematopoietic Stem Cells from cord blood CD133+ 100,000 cells 2C-102A HPGM PT-3926 CD133+ Hematopoietic Stem Cells from cord blood CD133+ 500,000 cells 2C-102 HPGM PT-3926 CD34+ Hematopoietic Stem Cells from cord blood CD34+ 100,000 cells 2C-101B HPGM PT-3926 CD34+ Hematopoietic Stem Cells from cord blood CD34+ 500,000 cells 2C-101A HPGM PT-3926 CD34+ Hematopoietic Stem Cells from cord blood CD34+ 1 million cells 2C-101 HPGM PT-3926 MSC Human Mesenchymal Stem Cells PT-2501 MSCGM PT-3001 MSC Rat Mesenchymal Stem Cells PT-2505 Rat MSCGM 192853 NHOst Osteoblasts CC-2538 OGM CC-3207 NHAC-kn Articular chondrocytes, knee CC-2550 CDM CC-3225 HPdLF Peridontal ligament fibroblasts CC-7049 SCGM CC-3205 R-OST Rat calvariae osteoblasts R-Ost-583 DMEM high glucose and Rat MSCGM SQ kit N/A Osteoclalsts Osteoclast Precursors 2T-110 OCP PT-8001 CD133+ Bone Marrow CD133+ 100 000 cells 2M-102A HPGM PT-3926 CD133+ Bone Marrow CD133+ 500 000 cells 2M-102 HPGM PT-3926 Bone marrow Fresh Bone Marrow Cells on Demand N/A N/A Mammary epithelial CC-2551 MEGM CC-3150 Islets Fresh Human Pancreatic Islets 100K IEQ 201981 Supplemented Islet Media Included with Product Islets Fresh Human Pancreatic Islets 20K IEQ 201983 Supplemented Islet Media Included with Product Islets Fresh Human Pancreatic Islets 10K IEQ 210984 Supplemented Islet Media Included with Product Islets Fresh Human Pancreatic Islets 5K IEQ 201985 Supplemented Islet Media Included with Product Islets Fresh Human Pancreatic Islets 2K IEQ 202998 Supplemented Islet Media Islets Frozen Pancreatic Islets Cells on Demand N/A N/A Pancreas Acinar Tissue Cells on Demand N/A N/A Blood and Lymph Bone Breast HMEC Pancreatic Included with Product Continued on next page 17 BioResearch Cancer Research and Drug Discovery Tools Ordering Information Cell Type Description Cat# (cells) Recommended G rowth Media Media Bulletkit Cat # SAEC Small Airway epithelial CC-2547 SAGM CC-3118 NHBE with RA Bronchial/Tracheal epithelial with retinoic acid CC-2540 BEGM CC-3170 NHBE without RA Bronchial/Tracheal epithelial without retinoic acid CC-2541 BEGM CC-3170 NHLF Lung fibroblasts CC-2512 FGM-2 CC-3132 HMVEC-L Lung Microvascular Endothelial CC-2527 EGM-2MV CC-3202 HPAEC Pulmonary artery endothelial CC-2530 EGM-2 CC-3162 PASMC Pulmonary artery smooth muscle cells CC-2581 SmGM-2 CC-3182 BSMC Bronchial Smooth muscle CC-2576 SmGM-2 CC-3182 HMVEC-Lly Lung lymphatic microvascular endothelial CC-2814 EGM-2MV CC-3202 UtMVEC-Myo Uterine microvascular endothelial CC-2564 EGM-MV CC-3125 UtSMC Uterine smooth muscle CC-2562 SmGM-2 CC-3182 UASMC Umbilical artery smooth muscle CC-2579 SmGM-2 CC-3182 PrEC Prostate epithelial CC-2555 PrEGM CC-3166 PrSC Prostate stromal CC-2508 SCGM CC-3205 PrSMC Prostate smooth muscle CC-2587 SmGM-2 CC-3182 HSeC Sertoli Cells MM-HSE-2305 SeGM 191053 Lung Reproductive BdSMC Bladder Smooth Muscle CC-2533 SmGM-2 CC-3182 HMVEC-Bd Bladder microvascular endothelial CC-7016 EGM-2MV CC-3202 HMVEC-dAd Adult dermal microvascular endothelial CC-2543 EGM-2MV CC-3202 HMVEC-dBIAd Adult dermal blood microvascular endothelial CC-2811 EGM-2MV CC-3202 HMVEC-dBlNeo Neonatal dermal blood microvascular endothelial CC-2813 EGM-2MV CC-3202 HMVEC-dLyAd Adult dermal lymphatic microvascular endothelial CC-2810 EGM-2MV CC-3202 HMVEC-dNeo Neonatal dermal microvascular endothelial CC-2505 EGM-2MV CC-3202 HMVEC-dLyNeo Neonatal dermal lymphatic microvascular endothelial CC-2812 EGM-2MV CC-3202 NHDF-Ad Adult dermal fibroblasts CC-2511 FGM-2 CC-3132 NHDF-Neo Neonatal dermal fibroblasts CC-2509 FGM-2 CC-3132 NHEK-Ad Epidermal keratinocytes, Adult 192627 KGM-Gold 192060 NHEK-Neo Epidermal keratinocytes, Neonatal 192907 KGM-Gold 192060 NHEK-Neo Pooled Epidermal keratinocytes, neonatal pooled 192906 KGM-Gold 192060 NHEM-Neo Neonatal normal human epidermal melanocytes CC-2504 MGM-4 CC-3249 NHEM-Ad Adult normal human epidermal melanocytes CC-2586 MGM-4 + ET-3 Supplement CC-3249 + CC-4510 Preadipocytes Preadipocytes, subcutan 4 million cells PT-5001 PGM-2 PT-8002 Preadipocytes Preadipocytes, visceral 1 million cells PT-5005 PGM-2 PT-8002 Preadipocytes Preadipocytes, sub cutan 1 million cells PT-5020 PGM-2 PT-8002 ADSC Adipose-derived stem cells PT-5006 ADSC PT-4505 Dermal (Skin) 18 Cell Type Description Cat# (cells) Recommended G rowth Media Media Bulletkit Cat # Skeletal Muscle Tissue HSMM Skeletal Muscle myoblasts CC-2580 SkGM-2 CC-3245 SkMC Skeletal muscle cells CC-2561 SkGM CC-3160 NHA Astrocytes CC-2565 AGM CC-3186 R-Cx Rat Brain cortex neurons R-Cx-500 PNGM CC-4461 R-Hi Rat brain hippocampus neurons R-Hi-501 PNGM CC-4461 R-Cp Rat brain striatum neurons R-Cp-502 PNGM CC-4461 R-Drg Rat dorsal root ganglion neurons R-Drg-505 PNGM CC-4461 R-eDrg Embryonic rat dorsal root ganglion neurons R-eDrg-515 PNGM CC-4461 R-Cb Rat cerebellar neurons R-Cb-503 PNGM-A CC-4512 R-HTh rat brain hypothalamic neurons R-HTh-507 PNGM CC-4461 R-CxAs Rat brain cortex astrocytes R-CxAs-520 AGM CC-3186 R-HiAs Rat brain hippocampus astrocytes R-HiAs-521 AGM CC-3186 R-CpAs Rat brain striatum astrocytes R-CpAs-522 AGM CC-3186 R-AsM Rat brain Cx-Hi-Cp mix astrocytes R-AsM-530 AGM CC-3186 M-Cx Mouse CD1 brain cortex neurons M-Cx-400 PNGM CC-4461 M-Cp Mouse CD1 brain striatum neurons M-Cp-402 PNGM CC-4461 M-Cx Mouse C57 brain cortex neurons M-Cx-300 PNGM CC-4461 M-Cp Mouse C57 brain striatum neurons M-Cp-302 PNGM CC-4461 M-AsM Mouse CD1 brain mixed astrocytes M-AsM-430 AGM CC-3186 M-AsM Mouse CD57 brain mixed astrocytes M-AsM-330 AGM CC-3186 M-Hi Mouse brain hippocampus neurons M-Hi-401 PNGM CC-4461 Neural Cells Continued on next page 19 BioResearch Cancer Research and Drug Discovery Tools Ordering Information Description Cat. No. Nucleofector™ Devices 4D-Nucleofector™ Core Unit AAF-1002B – 4D-Nucleofector™ X Unit AAF-1002X Requires core unit to build complete system 4D-Nucleofector™ Y Unit AAF-1002Y Requires core unit to build complete system 4D-Nucleofector™ Guarantee, 2-year extension AWE-1002 Has to be purchased at the time the device is purchased 4D-Nucleofector™ Service Contract AWC-1001 Can be purchased at any time during the guarantee period 96-well Shutte™ add-on (including laptop) AAM-1001S Requires core and X unit to build complete system 96-well Shutte™ Guarantee, 2-year extension AWM-1002 Has to be purchased at the time the device is purchased 96-well Shutte™ Service Contract AWB-1001 Can be purchased at any time during the guarantee period HT Nucleofector™ System AAU-1001 – HT Nucleofector™ Installation and Training AWT-1001 HT Nucleofector™ Service Contract AWU-1001 Nucleofector™ 2b Device AAB-1001 – Nucleofector™ 2b Guarantee, 2-year extension AWD-2002 Has to be purchased at the time the device is purchased Nucleofector™ 2b Service Contract AWA-2001 Can be purchased at any time during the guarantee period Can be purchased at any time during the guarantee period 100 μl Nucleocuvette™ 20 μl Nucleocuvette™; 16-well Dipping Electrode 12 rxn 24 rxn 32 rxn 24 rxn P1 Primary Cell 4D-Nucleofector™ X Kit V4XP-1012 V4XP-1024 V4XP-1032 – P2 Primary Cell 4D-Nucleofector™ X Kit V4XP-2012 V4XP-2024 V4XP-2032 – P3 Primary Cell 4D-Nucleofector™ X Kit V4XP-3012 V4XP-3024 V4XP-3032 – P4 Primary Cell 4D-Nucleofector™ X Kit V4XP-4012 V4XP-4024 V4XP-4032 – P5 Primary Cell 4D-Nucleofector™ X Kit V4XP-5012 V4XP-5024 V4XP-5032 – Primary Cell Optimization 4D-Nucleofector™ X Kit – – V4XP-9096 (96 rxn) – AD1 4D-Nucleofector™ Y Kit (adherent) – – – V4YP-1A24 AD2 4D-Nucleofector™ Y Kit (adherent) – – – V4YP-2A24 Optimization 4D-Nucleofector™ Y Kit (adherent) – – – V4YP-9A48 SE Cell line 4D-Nucleofector™ X Kit V4XC-1012 V4XC-1024 V4XC-1032 – SF Cell line 4D-Nucleofector™ X Kit V4XC-2012 V4XC-2024 V4XC-2032 – SG Cell line 4D-Nucleofector™ X Kit V4XC-3012 V4XC-3024 V4XC-3032 – Cell Line Optimization 4D-Nucleofector™ X Kit – – V4XC-9096 (64 rxn) – 4D-Nucleofector™ Kits 20 μl Nucleocuvette™; 96-well 96 rxn 960 rxn P1 Primary Cell 96-well Nucleofector™ Kit V4SP-1096 V4SP-1960 P2 Primary Cell 96-well Nucleofector™ Kit V4SP-2096 V4SP-2960 P3 Primary Cell 96-well Nucleofector™ Kit V4SP-3096 V4SP-3960 P4 Primary Cell 96-well Nucleofector™ Kit V4SP-4096 V4SP-4960 P5 Primary Cell 96-well Nucleofector™ Kit V4SP-5096 V4SP-5960 Primary Cell Optimization 96-well Nucleofector™ Kit V4SP-9096 (160 rxn) – SE Cell line 96-well Nucleofector™ Kit V4SC-1096 V4SC-1960 SF Cell line 96-well Nucleofector™ Kit V4SC-2096 V4SC-2960 SG Cell line 96-well Nucleofector™ Kit V4SC-3096 V4SC-3960 Cell Line Optimization 96-well Nucleofector™ Kit V4SC-9096 – 96-well Shuttle™ Kits 20 20 μl Nucleocuvette™; 384-well 768 rxn 3840 rxn P1 Primary Cell 384-well Nucleofector™ Kit V5SP-1002 V5SP-1010 P2 Primary Cell 384-well Nucleofector™ Kit V5SP-2002 V5SP-2010 P3 Primary Cell 384-well Nucleofector™ Kit V5SP-3002 V5SP-3010 P4 Primary Cell 384-well Nucleofector™ Kit V5SP-4002 V5SP-4010 P5 Primary Cell 384-well Nucleofector™ Kit V5SP-5002 V5SP-5010 Primary Cell Optimization 384-well Nucleofector™ Kit V5SP-9001 (384 rxn) – SE Cell line 384-well Nucleofector™ Kit V5SC-1002 V5SC-1010 SF Cell line 384-well Nucleofector™ Kit V5SC-2002 V5SC-2010 SG Cell line 384-well Nucleofector™ Kit V5SC-3002 V5SC-3010 Cell Line Optimization 384-well Nucleofector™ Kit V5SC-9001 (384 rxn) – 384-well Nucleofector™ Kits 100 µl Aluminum Cuvette 10 rxn 25 rxn 4x25 rxn Human B Cell Nucleofector™ Kit VAPA-1001 VPA-1001 VVPA-1001 Human T Cell Nucleofector™ Kit VAPA-1002 VPA-1002 VVPA-1002 Human CD34+ Cell Nucleofector™ Kit VAPA-1003 VPA-1003 VVPA-1003 Human Dendritic Cell Nucleofector™ Kit VAPA-1004 VPA-1004 VVPA-1004 Human NK Cell Nucleofector™ Kit VAPA-1005 VPA-1005 VVPA-1005 Mouse T Cell Nucleofector™ Kit – VPA-1006 VVPA-1006 Human Monocyte Nucleofector™ Kit – VPA-1007 VVPA-1007 Human Macrophage Nucleofector™ Kit VAPA-1008 VPA-1008 VVPA-1008 Mouse Macrophage Nucleofector™ Kit VAPA-1009 VPA-1009 VVPA-1009 Mouse B Cell Nucleofector™ Kit VAPA-1010 VPA-1010 VVPA-1010 Mouse Dendritic Cell Nucleofector™ Kit VAPA-1011 VPA-1011 VVPA-1011 VAPF-1001 VPF-1001 VVPF-1001 VAPE-1002 VPE-1002 VVPE-1002 Nucleofector™ II/2b Kits Primary Blood Cells Primary Bone/Cartilage Cells Human Chondrocyte Nucleofector™ Kit Primary Cardiac Cells Rat Cardiomyocytes - Neonatal Nucleofector™ Kit Primary Dermal Cells Human Keratinocyte Nucleofector™ Kit VAPD-1002 VPD-1002 VVPD-1002 Normal Human Epidermal Melanocyte-Neo Nucleofector™ Kit VAPD-1003 VPD-1003 VVPD-1003 Human Coronary Artery Endothelial Cell Nucleofector™ Kit VAPB-1001 VPB-1001 VVPB-1001 Human Umbilical Vein Endothelial Cell Nucleofector™ Kit VAPB-1002 VPB-1002 VVPB-1002 Human Microvascular Endothelial Cell Lung Nucleofector™ Kit VAPB-1003 VPB-1003 VVPB-1003 Basic Nucleofector™ Kit for Mammalian Endothelial Cells VAPI-1001 VPI-1001 VVPI-1001 Primary Endothelial Cells Primary Epithelial Cells Normal Human Bronchial Epithelial Cell Nucleofector™ Kit VAPK-1001 VPK-1001 VVPK-1001 Basic Nucleofector™ Kit for Mammalian Epithelial Cells VAPI-1005 VPI-1005 VVPI-1005 Continued on next page 21 BioResearch Cancer Research and Drug Discovery Tools Ordering Information 100 µl Aluminum Cuvette 10 rxn 25 rxn 4x25 rxn Human Dermal Fibroblast Nucleofector™ Kit VAPD-1001 VVPD-1001 VPD-1001 Mouse Embryonic Fibroblast Nucleofector™ Kit 1 VAPD-1004 VPD-1004 VVPD-1004 Mouse Embryonic Fibroblast Nucleofector™ Kit 2 VAPD-1005 VPD-1005 VVPD-1005 Mouse Embryonic Fibroblast Starter Nucleofector™ Kit VPD-1006 – – Basic Nucleofector™ Kit for Mammalian Fibroblasts VAPI-1002 VPI-1002 VVPI-1002 VAPL-1004 VPL-1004 VVPL-1004 Mouse Neuron Nucleofector™ Kit VAPG-1001 VPG-1001 VVPG-1001 Chicken Neuron Nucleofector™ Kit VAPG-1002 VPG-1002 VVPG-1002 Rat Neuron Nucleofector™ Kit VAPG-1003 VPG-1003 VVPG-1003 Basic Nucleofector™ Kit for Mammalian Neurons VAPI-1003 VPI-1003 VVPI-1003 Basic Nucleofector™ Kit for Mammalian Glial Cells VAPI-1006 VPI-1006 VVPI-1006 Human Aortic Smooth Muscle Cell Nucleofector™ Kit VAPC-1001 VPC-1001 VVPC-1001 Basic Nucleofector™ Kit for Smooth Muscle Cells VAPI-1004 VPI-1004 VVPI-1004 Human CD34+ Cell Nucleofector™ Kit VAPA-1003 VPA-1003 VVPA-1003 Human Mesenchymal Stem Cell Nucleofector™ Kit VAPE-1001 VPE-1001 VVPE-1001 Human Stem Cell Nucleofector™ Kit 1 VAPH-5012 VPH-5012 VVPH-5012 Human Stem Cell Nucleofector™ Kit 2 VAPH-5022 VPH-5022 VVPH-5022 Human Stem Cell Starter Nucleofector™ Kit VPH-5002 (18 rxn) – – Mouse Embryonic Stem Cell Nucleofector™ Kit VPH-1001 VAPH-1001 VVPH-1001 Mouse Neural Stem Cell Nucleofector™ Kit VAPG-1004 VPG-1004 VVPG-1004 Rat Neural Stem Cell Nucleofector™ Kit VAPG-1005 VPG-1005 VVPG-1005 Cell Line Nucleofector™ Kit R VACA-1001 VCA-1001 VVCA-1001 Cell Line Nucleofector™ Kit T VACA-1002 VCA-1002 VVCA-1002 Cell Line Nucleofector™ Kit V VACA-1003 VCA-1003 VVCA-1003 Cell Line Nucleofector™ Kit C VACA-1004 VCA-1004 VVCA-1004 Cell Line Nucleofector™ Kit L VACA-1005 VCA-1005 VVCA-1005 Cell Line Optimization Nucleofector™ Kit VCO-1001N (18 rxn) – – cGMP Cell Line Nucleofector™ Kit L – VGA-1005 – cGMP Cell Line Nucleofector™ Kit V – VGA-1003 – Nucleofector™ II/2b Kits Primary Fibroblasts Primary Hepatocytes Mouse/Rat Hepatocyte Nucleofector™ Kit Primary Neural Cells Primary Smooth Muscle Cells Primary Stem Cells Cell Line Kits 22 Cat. No. Size Suited for VS1-00500 500 µl Primary Cell Kits (4D-Nucleofector™ System): P1, P2, P3, and P4 Cell Line Kits (4D-Nucleofector™ System): SE, SF and SG Primary Cell Kits (Nucleofector™ II/2b Device): To determine the kits compatible with Nucleofector™ PLUS-1 Supplement please refer to the table provided on our website: www.lonza.com/n-plus Cell Line Kits (Nucleofector™ II/2b Device): R, T, V, and L Nucleofector™ PLUS Supplements Nucleofector™ PLUS Supplement 1 Cat. No. Description Size MycoAlert™ PLUS Mycoplasma Detection Kit 10 tests BioAssay Kits LT07-701 LT07-703 30 tests LT07-705 50 tests LT07-710 100 tests LT07-518 MycoAlert™ Assay Control Set 10 tests LT07-221 ViaLight™ Plus Cell Proliferation and Cytotoxity BioAssay Kit 500 tests LT07-121 1,000 tests LT07-321 10,000 tests LT17-221 500 tests (with 5 white TC plates) LT17-517 ViaLight™ 100% Lysis Control Set (sold separately) 10 ml (200 tests) LT07-217 ToxiLight™ Non-destructive Cytotoxity BioAssay Kit 500 tests LT07-1117 1,000 tests LT07-117 500 tests (with 5 white TC plates) 193339 AdipoLyze™ Lipolysis Detection Kit 1 x 96-wells PT-7009 AdipoRed™ Adipogenesis Assay Reagent 5 × 4 ml PA-1000 OsteoAssay Bone Plate 1 × 96-wells PA-1500 OsteoLyse™ Bone Resorption Assay Kit 1 × 96-wells PA-1503 OsteoImage™ Bone Mineralization Assay 5 × 96-wells 23 www.lonza.com /research www.lonza.com/cancer Contact Information North America Customer Service: 800 638 8174 (toll free) [email protected] Scientific Support: 800 521 0390 (toll free) [email protected] Europe Customer Service: +32 87 321 611 [email protected] Scientific Support: +32 87 321 611 [email protected] International Contact your local Lonza distributor Customer Service: +1 301 898 7025 Fax: +1 301 845 8291 [email protected] International Offices Australia Austria Belgium Brazil Denmark France Germany India Ireland Italy Japan Luxemburg Norway Poland Singapore Spain Sweden Switzerland The Netherlands United Kingdom + 61 3 9550 0883 0800 201 538 (toll free) + 32 87 321 611 + 55 11 2069 8800 808 83 159 (toll free) 0800 91 19 81 (toll free) 0800 182 52 87 (toll free) +91 40 4123 4000 1 800 654 253 (toll free) 800 789 888 (toll free) + 81 3 6264 0660 +32 87 321 611 800 16 557 (toll free) + 48 781 120 300 + 65 6521 4379 900 963 298 (toll free) 020 790 220 (toll free) 0800 83 86 20 (toll free) 0800 022 4525 (toll free) 0808 234 97 88 (toll free) Lonza Cologne GmbH 50829 Cologne, Germany For research use only. Not for use in diagnostic procedures. The Nucleofector™ Technology is covered by patent and/ or patent pending rights owned by the Lonza Group Ltd or its affiliates. Unless otherwise noted, all trademarks herein are marks of the Lonza Group Ltd or its affiliates. The information contained herein is believed to be correct and corresponds to the latest state of scientific and technical knowledge. However, no warranty is made, either expressed or implied, regarding its accuracy or the results to be obtained from the use of such information and no warranty is expressed or implied concerning the use of these products. The buyer assumes all risks of use and/or handling. Any user must make his own determination and satisfy himself that the products supplied by Lonza Group Ltd or its affiliates and the information and recommendations given by Lonza Group Ltd or its affiliates are (i) suitable for intended process or purpose, (ii) in compliance with environmental, health and safety regulations, and (iii) will not infringe any third party’s intellectual property rights. © 2014 Lonza Cologne GmbH. All rights reserved. CD-BR025 01/14 CD-BR025
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