请问各位大神有没有比较详细的mESC分化protocol？

prince

最近在培养mESC，之前一直检测增殖，但是现在要检测分化，小弟没做过啊，请问哪位大神可以提供一份比较详细的protocol？最好也有分化后的检测方法，有文献也可以推荐啊，谢谢啦！

ceshishua

首先你要说一下分化成什么细胞啊？
" M' Y0 d. V  n* ?4 t& t推荐一本书：http://product.dangdang.com/21066471.html
  c$ q$ V5 Y% h! x" [' y1 e. s《精编干细胞实验方法》
8 [. m9 t1 M3 m* T0 m7 p: a3 t8 L作     者（美）兰扎　等编* I5 c: Q6 M2 X! `, Q$ k3 K
出 版 社科学出版社; ]" r6 Q- l+ J9 `9 z
出版时间2011-3-1
& S. q! t0 ^) I6 N: m# a1 jＩＳＢＮ9787030301826

prince

就是简单的ES细胞向3胚层分化，然后用RT检测各胚层的表达mraker？

realyy10

去feeder,去LIF基本就行了，密度低一些。可以考虑做悬滴形成EB后再放到板里。

yuanyecat

# w- W  H2 w2 p9 S6 s' d0 R3 C

7 o8 F, g* H9 T2 X有一本书，叫《Current Protocols in Stem Cell Biology》，里面的charpter 1，section D 有你感兴趣的内容
0 [6 ?, h! W6 R! Z! |9 W% {3 v1D Germ Layer Induction/Differentiation of Embryonic Stem Cells
8 z8 b8 @0 F$ m% _0 b; Q1. Unit 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap3 U' i& B& N( h! u% k. x" a4 O2 u
2. Unit 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by
/ L% I  e5 u! E+ ~  \# v/ KSuspension and Hanging Drop Cultures
0 V9 j% q% S4 B% a3. Unit 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies% \& y- I) Q/ I: P
and a Description of the Hematopoietic Blast Colony Forming Assay7 k0 X- v: ]8 L* E* b4 F. W5 Y8 Y, a
4. Unit 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically
, c% u- {- B8 @8 @& n0 SDefined Culture Conditions
; }2 k) W5 B2 l; s- A- h5. Unit 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells

yuanyecat

还有一篇是讲非人类的胚胎干细胞分离，也是很不错的。‘Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells’: I. _1 g- k6 e0 g
http://onlinelibrary.wiley.com/d ... Authenticated=false