求干细胞定向诱导分化心肌细胞的鉴定方法

819zhang

' \- l/ j5 F: _8 t; X+ L4 b6 f

$ h5 H( V/ T0 }7 \' F求干细胞定向诱导分化心肌细胞的鉴定方法

splinkerette

Cardiac differentiation using either the growth factor– or the small
& _2 W$ J' h+ V0 Omolecule–based differentiation protocols proceeds rapidly. A rela-6 ^- U6 ]7 K' v: o' H8 k' ?/ h3 b
tively pure ( >95%) population of brachyury-expressing mesen-+ T) w" a3 W, W9 v
doderm cells can be detected after 1 d of differentiation. Gene
) x0 e6 X0 F  i) y6 ^3 M4 a4 }expression of the cardiac transcription factors NKX2-5 (ref. 19)
" C6 m* e! a; n8 T4 r1 nand ISL1 (refs. 20 and 21) begins at day 4, with the protein detect-; t/ f: f' o" K
able at day 5 (Fig. 2b). Cardiac troponin T (cTnT) can be readily
5 i- |) K+ _4 k# ?# |# U5 e1 tdetected at day 8 of differentiation. The presence of cardiomyocytes   J# s# |. M* x3 `- J( g7 p
can be easily established by visual observation of spontaneously   * q& W* e) I4 }# I7 I1 f! s
contracting regions. The first beating cluster of cells can be observed
' d- ?1 b* U! w9 U3 o, sbetween days 8 and day 10, depending on the individual cell line & T$ a$ U: F/ n; [& C
used. Robust spontaneous contraction occurs by day 12. Cardiac
. O3 ~1 o4 @' M% @2 `% Jmarker protein expression after onset of contractions can also be ) _, i7 ]  Y: g% o0 s7 h) e
assessed by immunostaining or be quantified with flow cytometry.

wf78365421

请问是胚胎干细胞还是间充质干细胞诱导心肌细胞？

splinkerette

回复 wf78365421 的帖子
6 l3 [' R; E1 i; j( F2 l6 g# ^: d. u$ y# i! a" @& K
反正问的是鉴定方法，无所谓哪种途径，只要是检测心肌细胞的marker就ok吧

Jiangshi90

染色或流式 TNNT2, alpha-actinin