请问ips培养中Y-27632的作用是什么

基因疗法

看ips培养中，在细胞复苏的时候要加Y-27632，之后的再换液就不用加了，请Y-27632的作用是什么？为什么只在ips细胞复苏的时候加

LingG

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Y-27632为ROCK inhibitor,防止细胞凋亡的。1 K: b2 g. ?: B+ X

$ y: q2 u' J/ S% g+ ~Rho激酶（Rho associated kinase, ROCK）,参与细胞信号通路。ROCK inhibitor为ROCK 抑制剂。: g1 f' K1 s) V3 y% [8 |
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关于ROCKi的详细介绍，给你找了一篇文文献。A ROCK inhibitor permits survival of dissociated human embryonic stem cells. " U- z2 z- ~. O1 _" F5 ]& w2 L4 @

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基因疗法

LingG 发表于 2017-3-31 10:56 - m1 W. @! i/ n8 N) @% ~7 g
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: k- Z9 b% ?# N' A7 z* H6 iY-27632为ROCK inhibitor,防止细胞凋亡的。

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太感激了

基因疗法

怎么给分啊？想给楼上网友加个分

基因疗法

LingG 发表于 2017-3-31 10:56
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  u) Z0 J5 t- t% ~3 s; RY-27632为ROCK inhibitor,防止细胞凋亡的。

# w, E, V! G+ z+ Q% T7 l1 P能再问一下，为什么Y-27632要在复苏细胞时加入，而之后的换液就不需要了呐？

LingG

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# w- s( c0 }& {这个我也不是很知道啦，不好意思。问了下同事，说的是：因为刚开始，还没有活化，ROCK容易启动，导致凋亡。等细胞长起来了，就可加可不加了。
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LingG

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" p$ ^8 ~. \3 Z% i0 X: \再来点英文的补充：
8 \. j3 f1 c0 _$ g7 C& u& p/ ZRho-associated protein kinase (ROCK) is a protein involved in the regulation of& B. h( p- ], S# @( c: Z$ ~
several forms of cellular behaviour. Among these is the regulation of apoptosis.
( l0 I: Y  a2 M0 C! }! T1 u& OWhen a cell is preparing to undergo apoptosis, caspase 3 will cleave the, `$ r0 R+ Z$ h0 b- t" G0 d: t8 e
autoinhibiting C-terminal of the ROCK protein and the cell will perform a
  H/ Z7 p; ^# w$ @( A; g0 {cytoskeletal change known as membrane blebbing. In order to prevent apoptosis
; ?7 W0 k7 L5 t* N/ p2 Y7 n% e% kROCK inhibitors (ROCKi) are added to stem cell cultures preventing the membrane2 \6 Z, B: q" q' g! u! a) @/ @! f& X
blebbing and increasing cell survival by 20-30%. The use of ROCKi is universal and  K5 x7 N4 V: J1 k/ z8 B9 Q
most scientists use this inhibitor to prevent cell death especially during single-cell
5 X; w9 U7 K0 d( I. ]passaging and clonal assays.
, @2 k: ^, g+ M( l) pBenefits of not using ROCKi
+ |% q/ f$ e# Y1 h0 H: ^4 [+ i( y• Less is always more when culturing stem cells, fewer components added is beneficial both from an economical and a functional standpoint.3 t8 i$ L! {- @; @) Z! J- T
• There is a concern throughout the cell culture community that the ROCKi might affect the morphology and genetic stability of the cells. ROCKi promotes the survival of “bad” cells that would otherwise have undergone apoptosis which could lead to complications. These claims have
2 A& ^: T' W5 Nnot yet been validated but studies are underway and we believe that this is a benefit even today." Q4 H7 q$ q2 e2 f' [+ n9 M

0 ]# h3 E$ b4 X; B其实现在已经有很多ROCKi-free的体系了:victory:

基因疗法

LingG 发表于 2017-4-1 13:48
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这个我也不是很知道啦，不好意思。问了下同事，说的是：因为刚开始，还没有活化，RO ...

- J9 w( I; H+ j$ r+ N' L$ s太感激指点了，多谢

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