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本帖最后由 细胞海洋 于 2012-2-29 00:52 编辑 1 V; \) s# p5 ^
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视频教程-人胶质母细胞瘤神经球培养方法
7 X& i8 b) O2 I0 D1 A C) MVideo Article0 s- y, A8 d1 V$ m- x/ |
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells
- ]' Z+ R% C. {2 x* }. ~Using the Neurosphere Assay8 t) D- w8 a7 R" |! K* w
Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds1: A& o1 l8 A' }- |
1Department of Neurosurgery, University of Florida# w7 P O7 M; T4 O- b
2Department of Anatomical Sciences, Shiraz University of Medical Sciences* T+ F5 ^( K, M# j0 t5 s7 N
Correspondence to: Hassan Azari at Hassan.azari@neurosurgery.ufl.edu
0 g% L; g- N# x! H& x% T( \$ ]URL: http://www.jove.com/video/3633/( C/ d: k% `8 M, p1 x& A# ^; H, L
DOI: 10.3791/3633. `( }+ m1 q% j+ l
Keywords: Neuroscience, Issue 56, Glioblastoma Multiforme, Tumor Cell, Neurosphere Assay, Isolation, Expansion,5 x% K9 _, ^8 X" A k" ]
Date Published: 10/30/2011, W; z% Q) ?+ |2 P1 @
Citation: Azari, H., Millette, S., Ansari, S., Rahman, M., Deleyrolle, L.P., Reynolds, B.A. Isolation and Expansion of Human Glioblastoma Multiforme
( I$ y! }1 L5 `" NTumor Cells Using the Neurosphere Assay. J. Vis. Exp. (56), e3633, DOI : 10.3791/3633 (2011).: x2 z7 H& h# }7 |, L7 W
Abstract- H5 p! [1 Z5 N' h. K( j, S
Stem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in
" A. o. v1 h3 P. p5 E0 M# l/ Gglioblastoma mutliforme (GBM), is to identify and isolate tumor initiating cell population(s) to investigate their role in tumor formation, progression,' z% }- h, c0 {* J
and recurrence. Understanding tumor initiating cell populations will provide clues to finding effective therapeutic approaches for these tumors. The
* O3 N" ~$ v0 d' Eneurosphere assay (NSA) due to its simplicity and reproducibility has been used as the method of choice for isolation and propagation of many of; a1 U4 F3 q5 ~* o8 y( s
this tumor cells. This protocol demonstrates the neurosphere culture method to isolate and expand stem-like cells in surgically resected human
/ z1 q, M* O+ Y( |* Y7 UGBM tumor tissue. The procedures include an initial chemical digestion and mechanical dissociation of tumor tissue, and subsequently plating the4 R K- x! z. y! [$ p6 x7 }
resulting single cell suspension in NSA culture. After 7-10 days, primary neurospheres of 150-200 μm in diameter can be observed and are ready5 w0 T; x a1 P
for further passaging and expansion.
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发表于 2012-2-28 21:11
