A Manual for Primary Human Cell Culture
This manual is designed to serve as a practical guide to primary human cell culture, which is integral in both academic and industrial biotechnology research. As in the first edition, the content of the manual is not exhaustive, but rather contains selected protocols for specific cell types from major tissue groupings in the body. This improved second edition also includes a new section on stem cells and additional material on transfection. It should serve as a foundation for individual researchers to experiment, explore, and establish niche protocols for their specific needs. With its compact physical format that makes it portable and flexible for usage in a laboratory setting, the manual will be a useful guide for all beginners in primary human cell culture work.
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30 mins 3H]–thymidine AECs alamarBlue Amphotericin antibodies assay biosafety cabinet bone marrow buffer cell culture cell number cell pellet cell suspension cell types Centrifuge at 200 collagenase complete media concentration containing cryopreservation cryovials culture 100 culture flasks denaturation differentiation diluted dispase dissociation DMEM DMSO EDTA endothelial cells enzymatic digestion enzyme epithelial cells ethanol explant explant culture fibroblasts filter fluorescent growth haemocytometer Ham’s hESCs human BMSCs Immunocytochemistry incubator isolated Keratinocyte lentivirus manual medium membrane Mesenchymal method mg/ml microscope mins at room Mitomycin number of cells optimisation osteoblasts osteocyte Penicillin Penicillin 100 U/ml)–Streptomycin PicoGreen plasmid plate primary cell primary human cells protein protocols reagent Remove Requirements Resuspend Rinse room temperature Schantz serum serum-free media solution specific staining sterile Streptomycin supernatant supplemented techniques tissue sample transfection transfer trypan blue trypsin trypsin–EDTA U/ml U/ml)–Streptomycin 100 µg/ml Wash µg/ml µm cell strainer