![Rank: 1](static/image/common/star_level1.gif)
- 积分
- 25
- 威望
- 25
- 包包
- 145
|
回复 Demon_CN 的帖子
6 n" Y% R0 D6 R2 `2 y* r" h& [5 v# f# B. Q
Root apical papilla was gently separated from the surface of the root, minced and digested in! a3 F4 M- @( e$ a4 ~# b: h) ]
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and
: }9 S2 u6 |+ c* f+ I4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30& f4 I+ K8 M( Y2 s5 f: x5 t
minutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm' I- r8 ~1 x9 i) ~7 e+ P: n7 o( t
strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
! ? G& H. ?! D* w+ F3 {(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/
4 a1 I% b' t3 ?6 B9 C7 tInvitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-! H* _/ L. `/ ]% C( Q
Bio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM
$ i8 G0 L4 l6 o6 W+ b. Z: @L-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°5 I1 Z4 G; g0 i* D; _
C in 5% CO2. 这是我要养的这个间充质干细胞的培养方法,我想问的是:1:a-MEM和DMEM的区别,当然除了成分上的区别,是不是一个促进细胞的分化?有的人养牙髓干细胞会用GIBCO DMEM/F12(1:1) 11330,有的用hyclone的SH30265.01B。2:100 mM L-ascorbic acid 2-phosphate 这个L-抗坏血酸2-磷酸酯的具体的作用是什么呢?提供vc的营养,只是比vc稳定?可是矿化诱导液里面是加了抗坏血酸的,这不是促进干细胞的分化嘛?3:我按照文献里,查到http://www.wako-chem.co.jp/english/labchem/productinf.htm 这个公司的产品只能查到L-Ascorbic Acid Phosphate Mg Salt.nH2O。sigam公司也只能查到A8960 SIGMA L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate 。我到底该怎么办呢?这个细胞的培养 我到底该选择什么样的培养基,这个L-抗坏血酸2-磷酸酯 我该怎么买呢? |
|