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


CONCLUSIONS
These findings may contribute to a better understanding of the process of gliomagenesis.
Instituto do Cérebro, Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.
Lorena Favaro
LF
Pavon
Luciana Cavalheiro
LC
Marti
Tatiana Tais
TT
Sibov
Liza Aya Mabuchi
LA
Miyaki
Suzana Maria Fleury
SM
Malheiros
Javier Bustamante
JB
Mamani
Reynaldo Andre
RA
Brandt
Guilherme Carvalhal
GC
Ribas
Jorge Roberto
JR
Pagura
Marcos Augusto Stavale
MA
Joaquim
Hallin
H
Feres Junior
Lionel Fernel
LF
Gamarra
D000071916:AC133 Antigen; D015703:Antigens, CD; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002469:Cell Separation; D005909:Glioblastoma / Q000276:immunology / Q000473:pathology*; D006023:Glycoproteins; D006801:Humans; D007150:Immunohistochemistry; D014411:Neoplastic Stem Cells / Q000276:immunology / Q000473:pathology*; D064231:Nestin / Q000276:immunology*; D010455:Peptides
D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D064231:Nestin; C000606023:PROM1 protein, human; D010455:Peptides