Abstract
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. 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. 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. These findings may contribute to a better understanding of the process of gliomagenesis.
Highlights
According to the World Health Organization (WHO)(1), central nervous system (CNS) tumors are classified into seven major groups which include primary tumors and secondary tumors or metastatic
Glioblastoma tissue samples were used for establishing the primary culture of those tumors, which was homogeneous with spindle-shaped cells arranged in multidirectional bundles (Figure 1D)
Immunocytochemical tests revealed the expression of the antigenic marker CD133 in the tumor neurospheres of the glioblastoma primary culture (Figures 1D and F)
Summary
According to the World Health Organization (WHO)(1), central nervous system (CNS) tumors are classified into seven major groups which include primary tumors (neuroepithelial, meninges, paraspinal and cranial nerves, germ cell, sellar region, and hematopoietic) and secondary tumors or metastatic.Gliomas are tumors that arise from glial cells and include astrocytomas, oligodendrogliomas, oligoastrocitomas ( called mixed gliomas), and einstein. 2012;10(2):197-202198 Pavon LF, Marti LC, Sibov TT, Miyaki LA, Malheiros SM, Mamani JB, Brandt RA, Ribas GC, Pagura JR, Joaquim MA, Feres Junior H, Gamarra LF ependymomas. Glioma is described as the most common human CNS neoplasm,(2) and it is very difficult to treat due to several factors: (i) it is infiltrative; (ii) it is composed of cells with different morphological and functional characteristics expressing various neuronal markers; (iii) it is highly resistant to radiation and chemotherapy processes[5]. These characteristics are probably due to the competence of tumor cells which is similar to the competence of stem cells, explaining high rate of recurrence of the disease or its primary resistance to treatment. It is extremely important to search for new approaches referring to the genesis, progression and clinical behavior of brain tumors
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