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Root apical papilla was gently separated from the surface of the root, minced and digested in" D: R6 m( ]" P" I
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and. U, {$ g2 K% c& J8 |
4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30
) c# f1 @3 J9 P2 S% i) \minutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm5 r' ^6 D5 N/ s
strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
2 r/ ?- F' B8 s( ^' [4 _! v* w(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/( y1 f1 {' e$ p3 R
Invitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-
6 Q& K0 t( ^9 y% Z, eBio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM
! y2 M: |$ \$ n5 [, t! bL-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°
0 K' D6 r$ _8 A9 NC 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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