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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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视频教程-人胶质母细胞瘤神经球培养方法, }7 Z: `3 V/ I4 j
Video Article% ^, n5 ~# b8 w6 f( e* c
Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells" W5 E" j0 I+ L) p$ ] j
Using the Neurosphere Assay. o" Y3 F N% X0 }* p; u% f
Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds1
3 b1 [* ?$ F) B T1Department of Neurosurgery, University of Florida1 y' @2 t7 R' O) T9 F: l) c1 ~6 d3 B
2Department of Anatomical Sciences, Shiraz University of Medical Sciences
* b9 Y5 d3 {, r& `* y5 |* f; hCorrespondence to: Hassan Azari at Hassan.azari@neurosurgery.ufl.edu: z; e& I z$ ?: D) x6 ]& x0 d: D9 I1 v
URL: http://www.jove.com/video/3633/
' c! ^( T) m- R0 ~DOI: 10.3791/3633, q; n0 A8 w' w8 S; t
Keywords: Neuroscience, Issue 56, Glioblastoma Multiforme, Tumor Cell, Neurosphere Assay, Isolation, Expansion,
0 U* `& K f- R* N4 VDate Published: 10/30/2011
, f+ y# s# }2 ]( Q; t# wCitation: Azari, H., Millette, S., Ansari, S., Rahman, M., Deleyrolle, L.P., Reynolds, B.A. Isolation and Expansion of Human Glioblastoma Multiforme
( v) ] i$ D3 I4 jTumor Cells Using the Neurosphere Assay. J. Vis. Exp. (56), e3633, DOI : 10.3791/3633 (2011).) A: ?# U' e+ m' Q8 K
Abstract
$ t! G1 B8 O7 s1 [8 w0 n+ sStem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in+ I$ L& Y3 r" P" ~# Q
glioblastoma mutliforme (GBM), is to identify and isolate tumor initiating cell population(s) to investigate their role in tumor formation, progression,% F6 l1 D$ ~1 }+ ^
and recurrence. Understanding tumor initiating cell populations will provide clues to finding effective therapeutic approaches for these tumors. The
9 N+ ?3 u$ _( h1 ~' Q- ]6 Tneurosphere assay (NSA) due to its simplicity and reproducibility has been used as the method of choice for isolation and propagation of many of
/ {6 @1 E& v% ^& p/ W/ ~1 Xthis tumor cells. This protocol demonstrates the neurosphere culture method to isolate and expand stem-like cells in surgically resected human
9 l7 u; _& ]/ DGBM tumor tissue. The procedures include an initial chemical digestion and mechanical dissociation of tumor tissue, and subsequently plating the
! _, u6 g# y, ?( @! g/ d1 Kresulting single cell suspension in NSA culture. After 7-10 days, primary neurospheres of 150-200 μm in diameter can be observed and are ready# X3 |/ p9 ]. a2 f4 P
for further passaging and expansion.- r0 R H6 N3 J) n4 d; ]3 Q8 F' b
. ?7 _) @0 o/ F9 a* H2 c h4 S下载地址：包含PDF和视频
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发表于 2012-2-28 21:11
