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本帖最后由 细胞海洋 于 2012-2-29 00:52 编辑 - \6 A! Z% ^5 m! Q, j" O) K
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视频教程-人胶质母细胞瘤神经球培养方法. ]0 ^9 X, N( W$ y; v
Video Article: V0 e: d( X* x/ C; U
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells
3 u7 e, ~- b* t- {* v& T1 B( {Using the Neurosphere Assay
" F) O0 |& J5 s/ x, l: o, ` c3 _Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds15 _' E5 F7 h, h3 n e9 ~
1Department of Neurosurgery, University of Florida
, }' _- b: p/ |- O. }5 |2Department of Anatomical Sciences, Shiraz University of Medical Sciences# L) R; F5 ~) q# D) l3 V
Correspondence to: Hassan Azari at Hassan.azari@neurosurgery.ufl.edu5 `! a9 l1 d( r; c/ P' B+ \9 o, \
URL: http://www.jove.com/video/3633/) f, T' H0 I/ P, D, f' y8 E1 l' Q
DOI: 10.3791/3633
7 O' Q' L8 F1 v1 x4 B# A, F9 ]Keywords: Neuroscience, Issue 56, Glioblastoma Multiforme, Tumor Cell, Neurosphere Assay, Isolation, Expansion,
: B {( @& T! R/ o) c& T, q1 qDate Published: 10/30/2011( t+ K2 {- M& @$ G9 | c
Citation: Azari, H., Millette, S., Ansari, S., Rahman, M., Deleyrolle, L.P., Reynolds, B.A. Isolation and Expansion of Human Glioblastoma Multiforme" N0 S. N9 \0 f$ a: q) `4 b5 l
Tumor Cells Using the Neurosphere Assay. J. Vis. Exp. (56), e3633, DOI : 10.3791/3633 (2011)., d' F: E5 U$ G- ^9 o: G
Abstract
. e' ?) y' h# b& L$ OStem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in
# N2 Q3 ?& t, _/ e6 bglioblastoma mutliforme (GBM), is to identify and isolate tumor initiating cell population(s) to investigate their role in tumor formation, progression,
, r! n1 T3 N0 r/ z* G+ _( n, s, dand recurrence. Understanding tumor initiating cell populations will provide clues to finding effective therapeutic approaches for these tumors. The9 ]' y7 f$ h# P7 y3 V& h
neurosphere assay (NSA) due to its simplicity and reproducibility has been used as the method of choice for isolation and propagation of many of) t: C* A" [1 N6 X& a$ {# \# ^
this tumor cells. This protocol demonstrates the neurosphere culture method to isolate and expand stem-like cells in surgically resected human& m; u H) |- n' s& I, d
GBM tumor tissue. The procedures include an initial chemical digestion and mechanical dissociation of tumor tissue, and subsequently plating the
) O% A4 I" O9 S8 h1 d! b3 Fresulting single cell suspension in NSA culture. After 7-10 days, primary neurospheres of 150-200 μm in diameter can be observed and are ready2 P! ?' r5 _2 u! r
for further passaging and expansion. ~% F0 ]6 y9 {' F# L1 c4 x
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发表于 2012-2-28 21:11
