PMID- 22740855
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20211021
IS  - 1792-1074 (Print)
IS  - 1792-1082 (Electronic)
IS  - 1792-1074 (Linking)
VI  - 3
IP  - 1
DP  - 2012 Jan
TI  - A simplified and modified procedure to culture brain glioma stem cells from 
      clinical specimens.
PG  - 50-54
AB  - In recent years, the theory of brain glioma stem cells (BGSCs) has facilitated 
      the study of gliomas. BGSCs have been accepted as the origin of gliomas and 
      determine their biological features. Numerous efforts have been made to probe 
      into the biological characteristics and behaviors of BGSCs. However, the 
      culturing of target cells remains the essential first step for research on BGSCs. 
      In this study, we established a simplified procedure to culture and isolate BGSCs 
      from samples of clinical glioma patients. Samples of 17 glioma patients were 
      included in the study, and the processed glioma cells were grown in serum-free 
      stem cell media. After the tumor spheres appeared, a proliferation assay, a 
      single-cell-derived colonies formation assay and an induced differentiation assay 
      were carried out, followed by an immunocytochemistry assay. Serial passage was 
      used to purify the target cells, whereas neither animal experiments nor sorting 
      techniques were included. As a result, CD133(+) BGSCs from 8 out of 17 patients 
      were grown and maintained in a serum-free condition combined with EGF, FGF and 
      B-27 supplements. The tumor sphere cells were serially passaged and showed 
      pluripotency in an induced differentiation assay. Immunocytochemistry identified 
      the committed markers (CD133, GFAP and TU-20) and confirmed the cells were BGSCs 
      and their progeny. The results proved that CD133(+) BGSCs from resected glioma 
      tissue may be cultured in serum-free stem cell media, and may also be purified by 
      conditioned culture combining serial passage, which is time-saving and 
      cost-effective, and allows the cells to be used for subsequent research. The cell 
      sorting techniques and animal experiments of tumorigenecity are optional. Thus, 
      this modified procedure is more practical and feasible than other available 
      procedures.
FAU - Qiu, Bo
AU  - Qiu B
AD  - Department of Neurosurgery, First Hospital of China Medical University, Shenyang, 
      Liaoning 110001, P.R. China.
FAU - Zhang, Dongyong
AU  - Zhang D
FAU - Tao, Jun
AU  - Tao J
FAU - Wu, Anhua
AU  - Wu A
FAU - Wang, Yunjie
AU  - Wang Y
LA  - eng
PT  - Journal Article
DEP - 20111004
PL  - Greece
TA  - Oncol Lett
JT  - Oncology letters
JID - 101531236
PMC - PMC3362403
EDAT- 2012/06/29 06:00
MHDA- 2012/06/29 06:01
PMCR- 2011/10/04
CRDT- 2012/06/29 06:00
PHST- 2011/07/01 00:00 [received]
PHST- 2011/09/26 00:00 [accepted]
PHST- 2012/06/29 06:00 [entrez]
PHST- 2012/06/29 06:00 [pubmed]
PHST- 2012/06/29 06:01 [medline]
PHST- 2011/10/04 00:00 [pmc-release]
AID - ol-03-01-0050 [pii]
AID - 10.3892/ol.2011.433 [doi]
PST - ppublish
SO  - Oncol Lett. 2012 Jan;3(1):50-54. doi: 10.3892/ol.2011.433. Epub 2011 Oct 4.