[Isolation and culture of tumor stem cells from human brain glioma tissues]

Zhonghua Zhong Liu Za Zhi. 2006 May;28(5):331-3.
[Article in Chinese]

Abstract

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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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*
  • Neoplastic Stem Cells / metabolism
  • Peptides / metabolism

Substances

  • AC133 Antigen
  • Antigens, CD
  • Glial Fibrillary Acidic Protein
  • Glycoproteins
  • MAP2 protein, human
  • Microtubule-Associated Proteins
  • PROM1 protein, human
  • Peptides