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回复 基因疗法 的帖子) Q6 V4 M/ R+ K6 h& N L! J. c& k
; e) j2 P, s, w9 a, @3 b2 `) `再来点英文的补充:
$ _) S; U. V: a5 i9 Y5 k& LRho-associated protein kinase (ROCK) is a protein involved in the regulation of
, n3 N# o: K9 G+ s: G3 U% M; wseveral forms of cellular behaviour. Among these is the regulation of apoptosis.
' [. S9 z( m" h$ d" ~When a cell is preparing to undergo apoptosis, caspase 3 will cleave the0 V* |2 K# \, g, Z
autoinhibiting C-terminal of the ROCK protein and the cell will perform a$ \/ q/ E( r6 G/ P
cytoskeletal change known as membrane blebbing. In order to prevent apoptosis8 F. }2 ^$ B; g u5 \4 u+ [
ROCK inhibitors (ROCKi) are added to stem cell cultures preventing the membrane
2 x+ X o+ s7 a0 P3 q* s5 Zblebbing and increasing cell survival by 20-30%. The use of ROCKi is universal and( V0 u% _1 m% O- N$ w
most scientists use this inhibitor to prevent cell death especially during single-cell- ~8 }, ^* Z S/ h M! P
passaging and clonal assays. & s, U/ K( m1 A4 e
Benefits of not using ROCKi
, j2 O4 c% Y$ }5 A8 F! f• Less is always more when culturing stem cells, fewer components added is beneficial both from an economical and a functional standpoint.
1 Z3 V$ {: U4 ?! ?: z• 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 have4 v7 D4 r. ~, x$ f& H0 d6 a3 \* N" Y% J
not yet been validated but studies are underway and we believe that this is a benefit even today.
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其实现在已经有很多ROCKi-free的体系了:victory: |
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