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你好,那我想问,我再养人的牙根尖乳头干细胞。Root apical papilla was gently separated from the surface of the root, minced and digested in8 O* m% Q# m7 V. O" N5 ^
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and) W) Q' M+ r* K1 p5 C
4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30
; M! y5 [, c4 A. Q" k9 d) Zminutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm
. B- G: Q/ C- Q, p- ~strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
" q, P$ A) a y' M$ m. J(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/# @$ z) M* R# c# h8 B
Invitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-* U; n; h* ~. h
Bio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM) ~ O3 I( C: v5 I9 K, C% ~6 R V
L-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37° {/ r' i P, u! ]3 u/ G3 b
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?麻烦您了。 |
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