Isolation and culture of tumor stem cells from human brain glioma tissues.
- Author:
Qiang HUANG
1
;
Jun DONG
;
Yu-de ZHU
;
Quan-bin ZHANG
;
Xiao-yan JI
;
Ai-dong WANG
;
Qing LAN
Author Information
- Publication Type:Journal Article
- MeSH: AC133 Antigen; Antigens, CD; metabolism; Brain Neoplasms; pathology; Cell Differentiation; Cell Proliferation; Cell Separation; Cells, Cultured; Glial Fibrillary Acidic Protein; metabolism; Glioma; pathology; Glycoproteins; metabolism; Humans; Microtubule-Associated Proteins; metabolism; Neoplastic Stem Cells; cytology; metabolism; Peptides; metabolism
- From: Chinese Journal of Oncology 2006;28(5):331-333
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo isolate and culture tumor stem cells from glioma tissues obtained at surgical operation and to study their biological characteristics.
METHODSGlioma tissues obtained from surgically resected specimens of 8 patients were fully chopped, trypsinized, and filtered to prepare single cell suspensions. The cells were cultured in serum-free medium with EGF, LIF and bFGF. CD133(+) cells were purified by magnetic cell sorting, and cultured continuously in vitro to obtain tumor cell spheres. Tumor stem cells of the 5th passage were induced to differentiate with 10% FBS, and expression of cell differentiation markers such as Nestin, MAP2, GFAP was evaluated with immunocytochemistry techniques.
RESULTSCD133(+) cells were successfully separated and cultured from one anasplastic mixed astrocyte-ependymocyte type glioma specimen. These cells maintained a sphere-like growth status in vitro (3 months, 14 passages), and can self-renew, proliferate and conditionally differentiate into MAP2(+) and GFAP(+) cells. However, CD133(-) cells did not possess these properties.
CONCLUSIONGlioma tissue contains tumor stem cells. Those cells can be cultured and passaged in vitro for a long term, and therefore to offer new approaches for studying cellular and molecular biology of glioma.
