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Tumorsphere culture4 J3 C# F# o7 O# b! g1 d
The sorted MiaPaCa2 cells were suspended in serum-free: Y/ ^2 V8 R6 u2 W2 R; N
culture medium DMEM containing 1% N2 supplement, 2% B27
" c/ ~" A+ F( |, G# [3 vsupplement, 1% antibotic-antimycotic (Invitrogen), 20 ng/ml( v2 t* w+ K Q" f9 {' l3 a, A; T
human FGF-2 (Sigma), and 100 ng/ml EGF (Invitrogen), and
. k G( y; c1 y7 wplated in 24-well ultra-low attachment plates (Corning) 2,000 cells
& y. l9 N* l% sper well. 7–10 days later, plates were analyzed for tumorsphere R+ V: ~( G) B6 U8 \; S5 t% B
formation and were quantified using an inverted microscope3 \3 Z% a0 a# m$ o F7 F* M
(Olympus) at 100X, 200X, and 4006 magnifications. For8 [* o* S$ m: @3 K4 }
subsequent quantification of cell numbers per tumorsphere, a' b/ K" d' s, U( _5 E
tumorspheres were collected with a 40 mm sieve (BD Biosciences,San Jose, CA) and disassociated with trypsin to make a single cell1 c+ f( T4 X1 M# I$ p# W$ y# T3 y" o
suspension. The viable cells were then counted using trypan blue7 {( G. a$ g, ?
exclusion.# l7 D# [9 j3 Q" j
paper title:MicroRNA miR-34 Inhibits Human Pancreatic Cancer
6 |# M, N$ ]4 a; M V) v* wTumor-Initiating Cells0 x! g" T" z- @5 R' [
大家看看有没参考意义！ |
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发表于 2010-3-3 09:45
发表于 2010-2-25 15:36
