Improved technique for establishing short term human brain tumor cultures.

摘要: Culturing human central nervous system tumors has been difficult compared to other neoplasms. We report improved success rates for establishing short term brain tumor cultures using a modified tissue processing technique. Eighty-seven specimens (56 glioblastomas, 8 mid grade astrocytomas, oligodendrogliomas, 15 other) were obtained from June 1988 March 1997. The first twenty-three samples processed by dissection, partial enzyme dissociation, and filtration through culture sieve. Subsequent identically except cells centrifuged on density gradient prior plating. Successful defined as those surviving greater than three passages in growing sufficient numbers (>106 cells) allow freezing. Success rate was 42% (10/23) standard methods 86% (55/64) with the addition of centrifugation. Glial fibrillary acidic protein (GFAP) vimentin staining, karyotypes, growth curves representative glioma cultures. All tested positive (29/29) while 62% (18/29) GFAP. Of four karyotyped (two two oligodendrogliomas), all but one oligodendroglioma exhibited clonal cytogenetic abnormalities. These immunohistochemical karyotypic results are consistent malignant glial origin these cells. note, low passage grew slower more contact inhibition immortalized glioblastoma cell lines. Nevertheless, this simple method should aid further developing pre-clinical models well enhancing clinical applications dependent vitro adjust.