Isolation and characterization of a human glioblastoma cell line

Abstract

The characterization of a glioblastoma cells line represents an important achievement in the field of brain tumor research activity, which enables the researchers to obtain strong results with a wide recognition by the scientific community. The objective of this study was to develop a standardized glioblastoma cells line using the resources obtained entirely from the national research projects. The tumor samples was initially obtained intraoperative from a 58 years-old female patient, with the preoperative written consent of the patient. The histopathological exam showed a grad IV WHO glioma (glioblastoma). The sample was prepared by manual fragmentation into pieces up to few millimeters each, followed by enzymatic digestions using different concentration of trypsin. Then, the cellular line was cultivated for more then 100 passages using Dulbecco Modified Eagle's medium enrich with 10% fetal bovine serum. The characterization of the glioblastoma line was made by the evaluation of cells proliferation (growth curve), by morphological studies, by studying the karyotype of the cells and by immunohistochemistry studies for the identification of the specific markers (glial fibrillary acidic proteins - GFAP, vimentin) of the tumor cells. The determination of the expression/amplification of the receptors tyrosine kinases (PDGFR and more important EGFR) is in progress. The characterization of a glioblastoma cells line represents an essential step in order to obtain a better in vitro and in vivo experimental model for glioblastoma.

Keywords

• glioblastoma
• cells line
• karyotype
• immunohistochemistry