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回复 Demon_CN 的帖子; L L3 l1 W8 O# q0 i; s
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Root apical papilla was gently separated from the surface of the root, minced and digested in
0 @! V0 }+ \* C, W3 ^a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and
p) ?( v+ C2 h! g4 `; G! ^! M4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30
0 V$ @+ t+ \& C/ k3 F1 Y5 N+ v! \2 |! sminutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm( I' h0 `( ]; o* D
strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
2 N9 c) |; ~) C( R7 Q(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/
1 x _ `8 E' U+ e# ?! sInvitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-% y2 f7 v$ T: R1 u1 v8 G
Bio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM
: [ Q& t- a, I0 |2 Q1 q( ^% FL-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°+ ]( T( v, e% l6 A
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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发表于 2012-3-21 17:10
发表于 2012-3-22 23:53
