Einstein (São Paulo) (Jun 2012)

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

  • Lorena Favaro Pavon,
  • Luciana Cavalheiro Marti,
  • Tatiana Tais Sibov,
  • Liza Aya Mabuchi Miyaki,
  • Suzana Maria Fleury Malheiros,
  • Javier Bustamante Mamani,
  • Reynaldo Andre Brandt,
  • Guilherme Carvalhal Ribas,
  • Jorge Roberto Pagura,
  • Marcos Augusto Stavale Joaquim,
  • Hallin Feres Junior,
  • Lionel Fernel Gamarra

Journal volume & issue
Vol. 10, no. 2
pp. 197 – 202

Abstract

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OBJECTIVE: To establish the method of isolation and culture of human glioblastoma neurospheres, and the purification of their stem cells, followed by the process of obtaining tumor subspheres, immunophenotypically characterizing this clonogenic set. METHODS: Through the processing of glioblastoma samples (n=3), the following strategy of action was adopted: (i) establish primary culture of glioblastoma; (ii) isolation and culture of tumor neurospheres; (iii) purify cells that initiate tumors (CD133+) by magnetic separation system (MACS); (iv) obtain tumor subspheres; (v) study the expression of the markers nestin, CD133, and GFAP. RESULTS: The study successfully described the process of isolation and culture of glioblastoma subspheres, which consist of a number of clonogenic cells immunophenotypically characterized as neural, which are able to initiate tumor formation. CONCLUSION: These findings may contribute to a better understanding of the process of gliomagenesis.

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