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回复 Demon_CN 的帖子3 O* C* R% O" x
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Root apical papilla was gently separated from the surface of the root, minced and digested in
. y+ S3 i$ P! {/ v, ^. n5 pa solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and/ ]: K) r/ b/ k$ V Z+ X
4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30* }8 `% Y6 j! L, x
minutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm
, i; V! S+ d8 ? X+ R8 K8 fstrainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
: S: ~, _( d$ S9 |+ Z(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/
& [8 g( [! H4 CInvitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-
. [ ^. P" z: J3 e; c+ C( L0 L$ jBio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM9 u# n: |; ^. _: @- p) A
L-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°
3 {5 V! w- E1 D, 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? |
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