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参考下这个吧1 Z$ n: o" t0 \" g+ d
from In Vitro Cell.Dev.Biol.—Animal (2009) 45:573–576
% i$ k7 |! d9 q+ ^3 [To isolate stem cells, umbilical cord samples
# s# G+ ^+ x$ C+ \" q; ]were rinsed in 75% ethanol (Sigma, Poole, UK) for 30 s
3 y8 C3 N* d ^( } [) \# \and cut open in parallel to umbilical cord vessels, so as; j( p" l# R; C, z- S
to expose them fully. The gelatinous tissue surrounding4 U( M( Y A6 J: u
the vessels was excised and minced into very fine pieces! B/ a- r1 N, Z9 v8 u
of 0.5–1 mm2, which were plated on a sterile 100×20 mm2 ~5 f8 w L" G6 l% }& `
petri dish (Corning, Ewloe, UK) and left for 5–10 min at
: w0 d b* P v* p2 z broom temperature, to facilitate tissue attachment. The
" q# [. g8 r4 L. k3 [! S+ A& p6 sminced tissue was carefully covered with 5 ml of growth
* K. c/ k* a' V1 u9 G( {- \medium comprised of low-glucose DMEM supplemented- I4 }' @+ j% A8 Q. e7 z
with 10% foetal bovine serum, 2 mM L-glutamine,
, S9 k6 M* n# I" b O/ ~& }4 T0 Ypenicillin (100 U/ml) and streptomycin (100 μg/ml) solution,9 ^6 ~( i5 y) f% Y7 p) ~) `
25 μg/ml Fungizone, 5 ng/ml basic fibroblast growth factor4 ] i; g3 O3 R+ H* e* A0 n( k
and 5 ng/ml epidermal growth factor (all from Invitrogen,
6 n# c/ A+ W3 m1 cPaisley, UK). WJ samples were incubated at 37°C in a
) L& t! Q( n) V! W5 nhumidified CO2 incubator for 5–10 d, before visible colonies p5 s! J( c E: n
of WJ HUMSCs were observed. |
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发表于 2010-5-3 11:02
发表于 2010-5-4 20:12
