参考下这个吧: j0 O) ]/ ?) v
from In Vitro Cell.Dev.Biol.—Animal (2009) 45:573–576 9 J) H2 }* @" ^. \& a" hTo isolate stem cells, umbilical cord samples8 M1 O$ r! h( Y) q" ?7 X6 x
were rinsed in 75% ethanol (Sigma, Poole, UK) for 30 s! C; G( O3 y7 K/ m8 A, [/ K
and cut open in parallel to umbilical cord vessels, so as0 T& r8 ` ?7 K& Y' d- S; V
to expose them fully. The gelatinous tissue surrounding8 W$ k8 [! ?( i$ {2 M
the vessels was excised and minced into very fine pieces ; D6 C+ a6 H. P5 Cof 0.5–1 mm2, which were plated on a sterile 100×20 mm! I8 U" k# ]+ ]! O6 G: E
petri dish (Corning, Ewloe, UK) and left for 5–10 min at& e9 w, v6 u. u: }1 o w; n& G
room temperature, to facilitate tissue attachment. The 5 r- R( F; I5 Y6 P4 Ominced tissue was carefully covered with 5 ml of growth1 B- d: `) B- s k4 r3 _+ {* ~
medium comprised of low-glucose DMEM supplemented 7 J" _4 k5 d: a) Z7 |& A% T8 s5 `with 10% foetal bovine serum, 2 mM L-glutamine, 2 @5 Z2 D+ L4 d/ v& R4 q. p% Ppenicillin (100 U/ml) and streptomycin (100 μg/ml) solution,5 |8 @$ b4 A5 J
25 μg/ml Fungizone, 5 ng/ml basic fibroblast growth factor- z- n) ^* s) w" k% l7 x* f
and 5 ng/ml epidermal growth factor (all from Invitrogen, ' s3 k# U/ v, S% Q# tPaisley, UK). WJ samples were incubated at 37°C in a M E' i! v: V
humidified CO2 incubator for 5–10 d, before visible colonies 7 C5 y, }( i2 I& B1 X, vof WJ HUMSCs were observed.作者: 香瓜瓜 时间: 2010-5-7 15:26
