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- @. E. P2 Z' c9 [0 r! T5 ]) n: |Root apical papilla was gently separated from the surface of the root, minced and digested in! ]+ V k4 w1 d2 @. O7 w
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and
! c( ~9 \( D0 j4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30; C1 t7 \( _$ \# i2 L8 Q
minutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm) p0 n7 W' \7 ]& H9 w5 ^
strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes
1 d$ v5 ?' Z5 d. A {(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/- N1 w2 S* D, `; ~
Invitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-/ a0 O: B" N$ h- G4 V2 R
Bio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM9 ` `) i' {5 ?; w9 M; ]
L-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°
) ^5 T' ]6 B% Z5 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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发表于 2012-3-21 17:10
发表于 2012-3-22 23:53
