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- L1 l+ Q' h# a, s7 [ ~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 in' U$ i2 D0 r: k3 [4 a
a solution of 3 mg/ml collagenase type I (Worthington Biochemicals Corp., Freehold, NJ) and8 L) ]9 B$ H1 ]
4 mg/ml dispase (Roche Diagnostic/Boehringer Mannheim Corp., Indianapolis, IN) for 30
" x% T1 \( P7 zminutes at 37°C. Single cell suspensions of SCAP were obtained by passing through a 70 μm" z2 \! L: F' R. ]2 T: V- O; S6 f0 W
strainer (Falcon, BD Labware, Franklin Lakes, NJ), seeded at 1×104 into 10-cm culture dishes$ ?4 v/ v6 X! S5 y) H
(Costar, Cambridge, MA), and cultured with α-Modification of Eagle’s Medium (GIBCO/
* j" B( A& }: ^) R2 v- |. z! EInvitrogen, Carlsbad, CA) supplemented with 15–20% fetal bovine serum (FBS) (Equitech-3 i) [4 y% k9 M( J4 O4 m- P
Bio Inc., Kerrville, TX), 100 mM L-ascorbic acid 2-phosphate (WAKO, Tokyo, Japan), 2 mM
- `- o3 B: O3 p( X% t& \L-glutamine (Biosource/Invitrogen), 100 U/ml penicillin and 100 mg/ml streptomycin at 37°5 V z! D/ x/ M# Y' j6 O# a5 e
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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发表于 2012-3-28 13:23
