[Isolation and culture of tumor stem cells from human brain glioma tissues].
OBJECTIVE
To isolate and culture tumor stem cells from glioma tissues obtained at surgical operation and to study their biological characteristics.


METHODS
Glioma 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.


RESULTS
CD133(+) 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.


CONCLUSIONS
Glioma 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.
Huang
Qiang
Q
Neurosurgery Department and Brain Tumor Research Laboratory, 2nd Affiliated Hospital of Suzhou University, Suzhou 215004, China. szqhdnah@pub.sz.jsinfo.net
Dong
Jun
J
Zhu
Yu-de
YD
Zhang
Quan-bin
QB
Ji
Xiao-yan
XY
Wang
Ai-dong
AD
Lan
Qing
Q
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D001932:Brain Neoplasms / Q000473:pathology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005910:Glioma / Q000473:pathology*; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D010455:Peptides / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; D005904:Glial Fibrillary Acidic Protein; D006023:Glycoproteins; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; C000606023:PROM1 protein, human; D010455:Peptides