Isolation, cultivation and characterization of CD133+ stem cells from human glioblastoma

Abstract

Objective: To establish the method of isolation and culture ofhuman glioblastoma neurospheres, and the purification of theirstem cells, followed by the process of obtaining tumor subspheres,immunophenotypically characterizing this clonogenic set. Methods:Through the processing of glioblastoma samples (n=3), the followingstrategy of action was adopted: (i) establish primary culture ofglioblastoma; (ii) isolation and culture of tumor neurospheres; (iii)purify cells that initiate tumors (CD133+) by magnetic separationsystem (MACS); (iv) obtain tumor subspheres; (v) study theexpression of the markers nestin, CD133, and GFAP. Results: Thestudy successfully described the process of isolation and culture ofglioblastoma subspheres, which consist of a number of clonogeniccells immunophenotypically characterized as neural, which areable to initiate tumor formation. Conclusion: These findings maycontribute to a better understanding of the process of gliomagenesis.

Keywords

• Glioblastoma
• Cell culture
• Neoplastic stem cells
• Antigens