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回复 Demon_CN 的帖子" `# C4 U0 M; S6 b1 Z
1 c: B" I7 T% x! NRoot apical papilla was gently separated from the surface of the root, minced and digested in; {( `; L* S) `8 D$ z v) V
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and
" g9 N4 p- s+ e( }1 J7 N4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30
" z. Y& j; \/ G* Jminutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm
' S0 [+ ^9 J0 t+ L7 ?. z% istrainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
1 {9 D5 x/ m+ ]) E0 \0 t, \(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/
( G0 Z9 Z) |( }) d/ b* yInvitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-
/ f: c" O6 E4 k6 X6 X: SBio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM
7 x$ a, b7 ^0 h; D8 mL-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°
/ c) \9 ~8 g4 p4 n- _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? |
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