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Development of a clonogenic cell assay for human brain tumors
Author(s) -
Rosenblum Mark L.,
Vasquez Dolores A.,
Hoshino Takao,
Wilson Charles B.
Publication year - 1978
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/1097-0142(197806)41:6<2305::aid-cncr2820410632>3.0.co;2-u
Subject(s) - clonogenic assay , pathology , anaplastic astrocytoma , glioma , ependymoma , astrocytoma , transplantation , biology , medulloblastoma , cell culture , medicine , cancer research , genetics
An in vitro colony formation assay has been developed for the evaluation of human brain tumor therapy. Single cells that are derived from biopsy specimens and form colonies containing more than 25 cells are defined as clonogenic. Tumor disaggregation is most efficient using 0.25% trypsin and the optimal culture conditions include 4 weeks of incubation at 37 C with 5% CO 2 in complete medium supplemented with 30% fetal calf serum and x‐irradiated “feeder cells.” Transplantation of cells harvested from colonies resulted in tumor formation in nude mice. In the past 1 1/2 years, we have grown colonies from each of 37 evaluable tumors. The mean colony forming efficiency (CFE) was 0.0643% for 13 malignant gliomas (glioblastomas and anaplastic astrocytomas), and 0.0165% for 8 astrocytomas (Kernohan grades I and II, fibrillary and protoplasmic types). Furthermore, the 5 largest CFE s occurred in malignant gliomas. This implies a greater clonogenic capacity for cells derived from these more aggressive tumors. Differences in CFE s were greater between malignant gliomas than among astrocytomas. In contrast, the mean CFE was 0.0018% for 3 specimens of “normal brain.” The CFE s of 6 meningiomas were consistantly large with a mean of 1.5% which corresponds with previous observations of rapid in vitro proliferation for this tumor type. Colonies also developed from 2 oligodendrogliomas, 2 medulloblastomas, an ependymoma, a choroid plexus papilloma, a subependymoma, and 3 metastatic neoplasms. No correlation was observed between colony morphology or cell type and parent tumor histology. The colony formation assay will permit evaluation of the clonogenic tumor cell population and should contribute to our knowledge of human brain tumors, towards the development of more effective treatment regimens.