Enzymatic-Mechanical Dissociation 分离效果怎么样？有人做过么？

西瓜太郎

2.4 Enzymatic-Mechanical Dissociation  A new method combining enzymatic digestion and mechanical dissociation was established.
- j& Q4 n4 i7 `0 N6 T) fIn brief, paste-like tissues were treated with 15ml enzyme mixture at 37o
% W) ^9 A$ |) M5 [. ?* DC for 30 minutes with gentle agitation (180rpm) to  lose  the collagen matrix. Enzymatic reaction was stopped 6 ^3 T2 Y' u  L6 N6 v' v. p- F
by adding  an  equal volume of MSC complete media.  Mixture were mechanically % b, _2 u( V- o) K0 d1 U& i  G
homogenised by using hand held cell homogeniser (Wiggen Hauser, Germany) at 9000 rpm 3 `# r3 l6 ^) t: C6 W
for 5 min  4  cycles. Mixture was then filtered by using 70µm and 40µm cell strainer to ) P: A, \# I4 u9 m
exclude  the excessive tissue. The cells were washed, counted and seeded into 25cm2
5 ]! z: Y2 v# g% S- Z  tissue culture flask in MSC complete media at 1x106cell/cm2
  Z0 @  A* R0 F0 ~% R! {3 m
6 L. T* F' W7 q, ?; C结果：Enzymatic-mechanical dissociation method can provide constantly high
8 F/ v- e' G6 x4 @; Rmononuclear cell yield (84.65±12.60 x 106cells/cm, n=6) compared to enzymatic digestion % }  f2 t1 w' ]4 h% j+ J
method (0.50±0.33 x 106 cells/cm, n=5) (Table 2). The successful rate of generation of MSC ! d2 m9 s" c# i+ X. _
using enzymatic-mechanical dissociation method are relatively higher (70%), followed  by ! F6 }1 Y% B8 O- {. [5 n, z7 h
explant method (25%) and enzymatic digestion (0%).  

西瓜太郎

不知道有大虾做过这个不？结果差别真的有这么大么

呼吸

我也想知道，脐带答案。