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- 积分
- 557
- 威望
- 557
- 包包
- 148
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Tumorsphere culture8 a& m9 k: F6 j3 [: Z3 L' d
The sorted MiaPaCa2 cells were suspended in serum-free
& g, r9 p: s" f6 t0 o# G0 hculture medium DMEM containing 1% N2 supplement, 2% B27' Y' }( Y0 p F0 V! e3 ]
supplement, 1% antibotic-antimycotic (Invitrogen), 20 ng/ml/ `& }8 a* C, ^( c) X) N+ v
human FGF-2 (Sigma), and 100 ng/ml EGF (Invitrogen), and5 c8 m" y& W. N- w6 t
plated in 24-well ultra-low attachment plates (Corning) 2,000 cells2 }" Y. Q" q! \! a. A1 D
per well. 7–10 days later, plates were analyzed for tumorsphere* S) ^( \& c* w9 v
formation and were quantified using an inverted microscope8 v0 x3 Y B5 h3 N
(Olympus) at 100X, 200X, and 4006 magnifications. For& q9 h2 z* K$ I, V# [+ _5 {4 o- I
subsequent quantification of cell numbers per tumorsphere," I" t( U+ Y! m# T4 S
tumorspheres were collected with a 40 mm sieve (BD Biosciences,San Jose, CA) and disassociated with trypsin to make a single cell* i' W; O1 p$ W$ G0 }5 C" [
suspension. The viable cells were then counted using trypan blue N6 s4 ^* H/ M# w4 n
exclusion.+ D; Q! |5 [3 v9 Z. q
paper title:MicroRNA miR-34 Inhibits Human Pancreatic Cancer3 ?& I# e! c* \( Y+ _7 z
Tumor-Initiating Cells
- n: p: }* Y) f+ g& ?% J大家看看有没参考意义！ |
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发表于 2010-2-24 10:49
发表于 2010-2-25 15:36
