回复 Dongqin11 的帖子 6 G# \# J# B% @5 o- n4 e e; Q, i3 H7 L0 b0 y& b& P. E
accutase要比0.05%胰酶温和，但比胶原酶要强得多，比dispase也强，accutase是将iPS克隆消化成单细胞，我们在制作EB时用，饲养层和无饲养层培养的iPS细胞传代都用胶原酶。作者: lchanlon 时间: 2012-9-22 10:10
if you culture ips cells in feeder, the collagenase IV is a good choice. But for feeder free, Accutase and dispase is not good. I suggest you use 0.5 mM EDTA in PBS. for 33mm dish, add 2ml of 0.5 mM EDTA, incubate at room temperature for 3-5 minutes, aspirate the EDTA. Then add 1 ml of medium using 1 ml pipet, pipet up and down to disassociate the ips cell and transfer into a new tube.( s0 I, b$ `5 R# _- j
^$ s7 O/ g8 D7 p+ a* ]补充内容 (2012-9-22 10:12): 9 z+ \$ C, ], S3 PIn our lab, it works well. this protocol from James A Thomson laboratory作者: sizhengliu 时间: 2012-9-23 08:45