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
4 ^* m9 V- Z8 p! H7 ]$ E- Bhttp://www.sciencedirect.com/science/article/pii/S1934590909001593
1 h8 G8 f5 ]: h1 e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
8 R9 s" |# G2 U" T9 u4 O# iCell culture: r4 |! ?# A  ]( X0 W( h2 S7 a/ V
OG2-MEFs were cultured on gelatin-coated dishes in normal MEF media: high-glucose D-MEM
2 `6 Z9 N4 X  [  ]- J, F(Invitrogen) with 10% FBS, 0.1 mM non-essential amino acids, and 2 mM L-glutamine. piPS" L3 ]% ^3 v1 b5 f
cells were cultured on irradiated CF1 MEFs with normal mESC growth media, which consist of2 k7 ^: r, _! X- P4 A2 m
Knockout DMEM (Invitrogen) supplemented with 20% KSR, 0.1 mM 2-ME, 2 mM L-glutamine,
% |. A: h1 G1 }2 k8 H  t0 O  I( q0.1 mM non-essential amino acids, and 1000 units/ml LIF (ESGRO, Chemicon International). The
+ F4 z/ j0 c# B! _/ C7 epiPS cells were passaged every 3 days as a single cell suspension using 0.05% trypsin/EDTA. D+ x( C% |1 S+ q1 G9 i
and seeded at 1.0x104 cells per cm2 for routine culture. For feeder-free culture, cells are grown* p$ m( x% }- S4 s! O& m
on gelatin-coated tissue culture dishes in chemically defined media, which consist of Knockout5 M5 \: P4 e, l8 T  @
DMEM supplemented with 1xN2, 1xB27, 0.1 mM 2-ME, 2 mM L-glutamine, 0.1 mM nonessential
( J* h6 \' n' _$ [amino acids, 50 μg/ml BSA fraction V (GIBCO), 103 units/ml LIF and 10 ng/ml BMP4
! q& Q" c; ~- q0 w(R&D).
( D- |( f& X  B+ x" n0 @Generation of piPS cells
: J/ d+ t3 ], e9 Z- ^# G3 x: a  GOG2-MEFs were seeded at 5x104 cells per well in a 6-well plate coated with gelatin in normal. f, Z3 }5 r4 L6 Q" X9 I! m
MEF media (DMEM supplemented with 10% FBS). On the next day, media was changed to the1 X% X/ x7 Z: o
protein transduction media, which were prepared by mixing the recombinant reprogramming
( s8 W/ B# y5 B5 Z) R9 r( O3 S: Pproteins at the final concentration of 8 μg/ml with regular mES cell growth medium8 _9 F* f$ ?. R( D3 g
supplemented with 1000 units/ml LIF. After overnight culture in the protein transduction media,
" ?, k; c; j0 ]8 u2 Jmedia was changed to normal mESC growth media, and cells were cultured for additional 36
( V+ l& _$ y3 V& T7 W9 yhours before repeating the same protein transduction cycle. Total four protein transduction
) T3 E: w1 \6 f: l1 m' n, K1 `5 _cycles were applied on the cells. After completing protein transduction, cells were then
) Z9 z) K$ }8 h4 g5 L* X- spassaged onto irradiated CF-1 MEF feeder cells at day 9 in normal mESC growth media. Media
* u4 @- Q/ |7 }: o3 E5 Qwere changed every 3-4 days until GFP+ colonies were observed around day 30-35. GFP+8 t! H4 L( T6 k: E
colonies were then passaged onto new irradiated MEF feeder cells in normal mESC growth
! e, S6 |9 u& i, {/ O( A- Tmedia, and stably maintained and expanded as piPS cells. Some colonies were further selected- `' C7 u7 C7 y2 H
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). # b- e* n1 S) m  [1 m5 a
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 6 M2 v. ]5 o7 ?: P
normal mESC growth media, which consist of Knockout DMEM (Invitrogen) supplemented with 20% KSR, 0.1 ...

% P4 z. V/ {" R你复制的是normal mESC growth media。
3 m0 @, u  g# Q: m" n; y5 z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 : S8 n7 {% z+ `3 [, }/ _
ES培养液里要加LIF的，我们实验室有人养了很长时间了。

6 N! I* ?2 P0 n1 D% k( ]; 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。