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本帖最后由 细胞海洋 于 2012-2-29 00:52 编辑
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/ A, u$ S2 F0 Z4 M视频教程-人胶质母细胞瘤神经球培养方法
9 u1 \) g. f D, q7 \Video Article9 Z8 I+ I t- j6 c8 t3 C
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells7 `( `0 Y8 G; V3 _+ [
Using the Neurosphere Assay) d& P& J/ r( b( R _, t
Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds1
2 S! a: }! q, g; H1Department of Neurosurgery, University of Florida
$ E. |% W w$ n$ `2Department of Anatomical Sciences, Shiraz University of Medical Sciences1 C1 _- Z/ n2 L
Correspondence to: Hassan Azari at Hassan.azari@neurosurgery.ufl.edu+ x( h/ L: W- T) R3 l
URL: http://www.jove.com/video/3633/
5 R0 ]! Q& ]* X1 p8 G; G \ FDOI: 10.3791/3633
) _( f1 ^! H/ H) _8 DKeywords: Neuroscience, Issue 56, Glioblastoma Multiforme, Tumor Cell, Neurosphere Assay, Isolation, Expansion,
* l( {& j0 A* O+ f* n5 Y4 k: eDate Published: 10/30/2011' B5 K9 ]) o8 S+ j* \
Citation: Azari, H., Millette, S., Ansari, S., Rahman, M., Deleyrolle, L.P., Reynolds, B.A. Isolation and Expansion of Human Glioblastoma Multiforme* |8 E7 }/ m5 G& S6 Q- t1 u8 o
Tumor Cells Using the Neurosphere Assay. J. Vis. Exp. (56), e3633, DOI : 10.3791/3633 (2011)./ c$ l; j3 W* d o# y
Abstract
4 n$ W+ j) |" H+ jStem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in
- b( ~7 X% J, Q' z+ E! M. K% zglioblastoma mutliforme (GBM), is to identify and isolate tumor initiating cell population(s) to investigate their role in tumor formation, progression,
5 }: H) z3 h$ E* ]) |$ h+ hand recurrence. Understanding tumor initiating cell populations will provide clues to finding effective therapeutic approaches for these tumors. The
8 t! r6 N8 x' c ?$ b" oneurosphere assay (NSA) due to its simplicity and reproducibility has been used as the method of choice for isolation and propagation of many of
; B" p0 f& X( O# c: K# c: s3 Jthis tumor cells. This protocol demonstrates the neurosphere culture method to isolate and expand stem-like cells in surgically resected human% k, V+ T; n! S7 F# S' {
GBM tumor tissue. The procedures include an initial chemical digestion and mechanical dissociation of tumor tissue, and subsequently plating the
( X/ g5 p z6 I& Qresulting single cell suspension in NSA culture. After 7-10 days, primary neurospheres of 150-200 μm in diameter can be observed and are ready
& V# q! W8 c1 k& C4 J" G5 x) zfor further passaging and expansion.* ?7 X; u0 H3 o) O
* C6 i; o; @7 i- r9 N8 X$ P& n下载地址：包含PDF和视频* _: M( N Z/ T6 d G( x8 U. ]
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发表于 2012-2-28 21:11
