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参考下这个吧
9 D* Q9 x7 ?& s3 a, ?* ^from In Vitro Cell.Dev.Biol.—Animal (2009) 45:573–576
6 @# ]8 V% V4 U3 k; z/ F# ETo isolate stem cells, umbilical cord samples* r/ Z {- I3 t0 `6 @; s7 b" ^1 H
were rinsed in 75% ethanol (Sigma, Poole, UK) for 30 s3 V1 d: p+ N2 f! ^0 A
and cut open in parallel to umbilical cord vessels, so as
8 a& Z% Q7 m7 Q3 X4 b: hto expose them fully. The gelatinous tissue surrounding+ W/ w; f7 H1 b2 ]0 T
the vessels was excised and minced into very fine pieces
. d. K( ~ R$ {9 Bof 0.5–1 mm2, which were plated on a sterile 100×20 mm$ p2 Z; U u3 I" M9 F4 u, H
petri dish (Corning, Ewloe, UK) and left for 5–10 min at
; s5 i |% S; r6 H w, B1 a* T8 jroom temperature, to facilitate tissue attachment. The
8 C2 l, v1 Y" K, u- t+ J1 jminced tissue was carefully covered with 5 ml of growth/ K& o* U4 P. H; E" w! h C
medium comprised of low-glucose DMEM supplemented, a3 z+ m# |, y
with 10% foetal bovine serum, 2 mM L-glutamine,
8 H5 L: W, k8 u/ C% p* c1 b$ b' Upenicillin (100 U/ml) and streptomycin (100 μg/ml) solution,1 O1 H4 v: ^: f0 }
25 μg/ml Fungizone, 5 ng/ml basic fibroblast growth factor
9 h# L g# A; c0 b- @and 5 ng/ml epidermal growth factor (all from Invitrogen,1 D; W$ r4 P/ y9 J, g
Paisley, UK). WJ samples were incubated at 37°C in a& V8 g' z( ^( I5 `# V
humidified CO2 incubator for 5–10 d, before visible colonies
; p4 y9 {9 V- E: \$ j oof WJ HUMSCs were observed. |
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发表于 2010-5-3 11:02
发表于 2010-5-4 20:12
