Although significant progresses were made in the field of molecular biology of malignant cerebral gliomas, the prognostic of these tumors continues to be reserved. One of the therapeutic failure reasons is the incomplete knowledge regarding the origin of these tumors and cells features, which in fact represent an obstacle in developing a cell and molecular therapy guided against malignant cells responsible for the tumor development and for the therapeutic resistance. Initiation and characterization of glioblastoma cell lines represents an essential step in order to obtain a better in vitro and in vivo experimental model for glioblastoma. We describe here a new glioblastoma line, named T11, which was successfully isolated in our laboratories starting with a tumor sample obtained intraoperative from a 58 years-old female patient. The histopathological evaluation showed a grad IV WHO glioma (glioblastoma). The sample was prepared by manual fragmentation, followed by enzymatic digestions using different concentration of trypsin. The cell line has been cultivated for more than 150 passages. The characterization of the glioblastoma line consisted in the evaluation of cells proliferation capacity (growth curve), morphological features, karyotyping and identification of specific markers. We found that T11 expressed specific markers for glial progenitors and astrocytes (glial fibrillary acidic protein-GFAP); oligodendrocites (A2B5; O4), and microglia (CD45, CD 11b). Cells were negative for neuronal lineage markers like beta3-tubulin and NCAM. In order to evaluate the differentiation grade of T11 cell line, the presence of stem cell markers (nestin, CD133) was explored. T11l cells expressed higher level of nestin and lower level of CD133 comparing with standard glioblastoma cell line U87. T11 cell line expressed VEGF and Bcl-2, but not EGFR and Mdrl and Bax. This new line has distinct and unique characteristics when compared with standard glioblastoma cell line (e.g., U87) and may become a new and useful in vitro model for glioblastoma.