Improved technique for establishing short term human brain tumor cultures

Culturing human central nervous system tumors has been difficult compared t o other neoplasms. We report improved success rates for establishing short term human brain tumor cultures using a modified tissue processing techniqu e. Eighty-seven brain tumor specimens (56 glioblastomas, 8 mid grade astroc ytomas, 8 oligodendrogliomas, 15 other) were obtained from June 1988 to Mar ch 1997. The first twenty-three samples were processed by dissection, parti al enzyme dissociation, and filtration through a tissue culture sieve. Subs equent samples were processed identically except tumor cells were centrifug ed on a density gradient prior to plating. Successful cultures were defined as those surviving greater than three passages in tissue culture and growi ng to sufficient numbers (> 10(6) cells) to allow freezing. Success rate wa s 42% (10/23) using standard processing methods and 86% (55/64) with the ad dition of density gradient centrifugation. Glial fibrillary acidic protein (GFAP) and vimentin staining, karyotypes, and growth curves were obtained f or representative glioma cultures. All cultures tested were positive for vi mentin (29/29) while 62% (18/29) were positive for GFAP. Of four cultures k aryotyped (two glioblastomas, two oligodendrogliomas), all but one oligoden droglioma culture exhibited clonal cytogenetic abnormalities. These immunoh istochemical and karyotypic results are consistent with the malignant glial origin of these cells. Of note, low passage human glioma cultures grew slo wer and exhibited more contact inhibition than immortalized human glioblast oma cell lines. Nevertheless, this simple method for establishing short ter m human brain tumor cultures should aid in further developing human brain t umor pre-clinical models as well as enhancing clinical applications depende nt on in vitro human brain tumor cell growth adjust.