小鼠脂肪干细胞传代后老化，原因？？？

pdongjian

张兴美5342

"发现用一定分量的生长因子和维他命可以防止脂肪干细胞老化和维持其原始性。现在我培养的脂肪干细胞可以传代到P20都没有问题"。能求教具体的数据不

SBCLPC

回复 chichao0000 的帖子3 H  u( L: M. d1 Q& e

( _' M- w+ F" ^: k7 W3 ]2 S加vc会有其他副作用吗

kakapo

6 n+ W( L0 o/ |& Q     Your protocol is ok, but you may try this. Initially, your cells should be health and had not get damaged by collagenae digestion. The digestion time should be shortened. Also, when the digestion is performed in PBS or HBSS, the stromal cells are in starvation. Lactated Ringer's buffer provides energy source for cells. This conditioned digestion is good for the cells surviving and avoid of cell programing senescence after isolation from tissue.
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3 o% X$ [) y3 z. R, Q$ p* o& ~    2，充分剪碎，用0.1%的一型胶原酶37度振荡消化1h。(0.5 mg/ml in lactated Ring's solution) type II collagnease digestion for 30 minutes)
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xiuyinx

学习了

辛西雅

回复 冠忠 的帖子( ?0 U( F) x8 `( T- @  e5 ?3 d
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脂肪干密度传的低了，时间长了都太容易老化，求教需要加那种因子和维他命啊

kakapo

脂肪干密度传的低了，时间长了都太容易老化1 x: o7 S! ?% |
I have to agree this view point. Now, I am going to set a co-culture system to expand mouse MSC. At present, I did not yet see positive result. As I get positive information, I will inform it in this forum.

gonglizhong

加生长因子，不就相当于给干细胞加了激素，促进干细胞生长老化？* A2 b" V- b  O% A2 H* I

屋檐上的乌鸦

个人意见，你原代的时候得检测下脂肪干细胞的真伪，还有在消化的时候时间过长对细胞有损伤！在说一点，你的培养基和血清都是有问题的！

wanghebin

回复 zerollx 的帖子7 r/ ?8 V" c! t' d
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你好，刚看了你的帖子。
) ^6 I6 Y3 ~! U0 _- }有一点值得注意的是胰酶消化的时间太长，时间太长会造成细胞损伤。+ a+ m. }) S: V' `- k5 O
另外培养液可以尝试使用不含血清的完全培养基替代。
: o  J: }2 S8 s" f2 Z3 ^希望对你有帮助。