我是新手，求hiPS向心肌细胞分化的protocol，和诱导hiPS成功率最高的方法和试剂

xiaoyudan1989

各位大侠，我手里有ES细胞，想先看看分化情况，之后要买试剂盒了，有没有protocol和试剂公司推荐？

chentian820

xiaoyudan1989

回复 chentian820 的帖子! Y6 Q3 t' K, T6 G' J8 P

# [& q3 R1 I* g8 T- M5 j" L非常感谢！

frankieisme

EB法:The undifferentiated hips cell colonies were cultured on a mitotically inactivated mouse embryonic fibroblast (MEF) feeder layer. The culture medium consisted of 80% knockout high-glucose glutamine-free DMEM with sodium pyruvate supplemented with 20% serum replacement, 1 mmol/L L-glutamine, 0.1 mmol/L mercaptoethanol, 4 ng/mL human recombinant basic fibroblast growth factor, and 1% nonessential amino acid stock. To induce differentiation, hiPS cells were dispersed into small clumps with collagenase IV ( 1 mg/mL  for 20 minutes). The cells were then transferred to plastic Petri dishes, where they aggregated to form embryoid bodies and were cultured in suspension for 10 days. The EBs were then plated on 0.1% gelatin-coated culture dishes and examined daily for the appearance of spontaneous contractions.2 X, s: r! U( b1 k9 @: y0 V
单层细胞法:iPS cells were washed in DMEM/F12 basal media and incubated in dispase for 20 minutes, after which cells were harvested, dispersed mechanically into clumps of 10-20 cells, and dispersed at ratios of 1:12 onto matrigel-coated plates with 2ml fresh mTeSR supplemented with Y27632. further feeds involved half medium changes every 48 h, without the need to further supplement with ROCK inhibitor. Activin A and BMP4 can be helpful.