3T3一L1传代需小牛血清理由:当细胞从快速分裂到汇合和接触抑制状态时,细胞经历脂肪样转化前脂肪。培养基中的高血清含量可提高脂肪积累[PubMed ID:4426090 ]
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37C to facilitate dispersal.
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning? T-75 flasks (catalog #430641) are recommended for subculturing this product.--该耗材国内应用较少,前期种子阶段培养推荐康宁-3799-96孔U型低耗材替代。
二甲基亚砜敏感,解冻后应立即去除冷冻保护剂,在9-10毫升完整的完全培养基溶液中进行缓慢离心(125×g,10分钟)