请问那篇蛋白诱导ips的文章里regular mES cell growth medium的配方是什么？

mofatuzi

这篇文章补充材料里提到的
& V" Q9 q$ _' l1 q3 d1 [4 ^6 w* Vhttp://www.sciencedirect.com/science/article/pii/S19345909090015935 [, X( x; k$ `4 `! _0 [! V
是不是normal mESC growth media再加LIF？

DA_huyang

占个沙发:lol

mofatuzi

我把内容放出来，请各位分析一下，谢谢
. _% j/ |' Z* S0 w" b; @Cell culture
& O) ?8 _7 w2 y$ MOG2-MEFs were cultured on gelatin-coated dishes in normal MEF media: high-glucose D-MEM/ K& N6 u& u) i! u
(Invitrogen) with 10% FBS, 0.1 mM non-essential amino acids, and 2 mM L-glutamine. piPS
1 N2 v. y% t8 x% Tcells were cultured on irradiated CF1 MEFs with normal mESC growth media, which consist of7 _( l* q! A6 c: \+ e
Knockout DMEM (Invitrogen) supplemented with 20% KSR, 0.1 mM 2-ME, 2 mM L-glutamine,  D" e( ~; }) R. ?" V  W9 l
0.1 mM non-essential amino acids, and 1000 units/ml LIF (ESGRO, Chemicon International). The
1 _% c% i3 G3 p7 C  QpiPS cells were passaged every 3 days as a single cell suspension using 0.05% trypsin/EDTA( s$ Z2 G+ v: F: L. S% Z& E
and seeded at 1.0x104 cells per cm2 for routine culture. For feeder-free culture, cells are grown
; s, b8 R4 w0 t! k& f8 Oon gelatin-coated tissue culture dishes in chemically defined media, which consist of Knockout% t0 o. Z) F( U4 L& T' t7 a
DMEM supplemented with 1xN2, 1xB27, 0.1 mM 2-ME, 2 mM L-glutamine, 0.1 mM nonessential7 l& w9 X7 e, c+ [1 x
amino acids, 50 μg/ml BSA fraction V (GIBCO), 103 units/ml LIF and 10 ng/ml BMP4" _$ \9 c! W$ l0 E
(R&D).1 U( w3 N1 H7 v7 G! S  M, ^
Generation of piPS cells% K; H4 I4 @9 h5 ~4 H  s
OG2-MEFs were seeded at 5x104 cells per well in a 6-well plate coated with gelatin in normal* N0 Z' ^6 X- c' W
MEF media (DMEM supplemented with 10% FBS). On the next day, media was changed to the" c/ z, T/ A5 c; p
protein transduction media, which were prepared by mixing the recombinant reprogramming+ n" {$ O7 I/ u# {, k
proteins at the final concentration of 8 μg/ml with regular mES cell growth medium
$ f, S4 h8 f5 Q, @$ m- J$ N, f3 gsupplemented with 1000 units/ml LIF. After overnight culture in the protein transduction media,5 @: g5 n# _1 U( p1 p$ F8 _
media was changed to normal mESC growth media, and cells were cultured for additional 36
; g/ w9 R& G, G( W2 u1 {hours before repeating the same protein transduction cycle. Total four protein transduction
! l! c% s1 H& @. g" f% C8 [cycles were applied on the cells. After completing protein transduction, cells were then
- j3 _! w/ I; b9 q6 ^passaged onto irradiated CF-1 MEF feeder cells at day 9 in normal mESC growth media. Media( ~# p# z5 A, v4 p% ]
were changed every 3-4 days until GFP+ colonies were observed around day 30-35. GFP+
- k/ o) S- i  L6 @8 \! N* ~" Pcolonies were then passaged onto new irradiated MEF feeder cells in normal mESC growth) t7 A9 \. W1 @5 u& ?- B: s! z
media, and stably maintained and expanded as piPS cells. Some colonies were further selected9 a5 H& ?/ w% X9 K+ F
and expanded in the presence of pluripotin (1 μM) or PD0325901 (1 μM).

mofatuzi

有知道的吗

song58

normal mESC growth media, which consist of Knockout DMEM (Invitrogen) supplemented with 20% KSR, 0.1 mM 2-ME, 2 mM L-glutamine, 0.1 mM non-essential amino acids, and 1000 units/ml LIF (ESGRO, Chemicon International).
. w( `# d0 ?5 x' M人家不是写的挺清楚的嘛

ophden

ES培养液里要加LIF的，我们实验室有人养了很长时间了。

mofatuzi

song58 发表于 2012-3-30 11:59 . h! \3 l5 p; `$ C, L1 |
normal mESC growth media, which consist of Knockout DMEM (Invitrogen) supplemented with 20% KSR, 0.1 ...

: W  |* \9 ?9 b; v5 u) z! O你复制的是normal mESC growth media。
$ w, u+ ~0 D8 n后面这个regular mES cell growth medium不知道什么意思，与normal是什么关系？

mofatuzi

ophden 发表于 2012-3-30 14:07 ( ]+ a3 ~5 c" p
ES培养液里要加LIF的，我们实验室有人养了很长时间了。

- w1 ^$ ^+ ~' r0 N8 v0 W我知道要加LIF。只是不知道regular mES cell growth medium和normal mESC growth media是什么关系。