Multicellular tumor spheroids from human gliomas maintained in organ culture

Abstract

Tumor tissue from seven human gliomas was maintained in long-term agar overlay culture as multicellular organotypic spheroids. Light microscopic and ultrastructural observation of the spheroids displayed morphological features similar to those of the original tumor tissue in vivo; in this respect they were different from spheroids obtained from permanent cell lines. The spheroids contained preserved vessels, connective tissue, and macrophages, revealing a close resemblance to the conditions in the original tumor. Flow cytometric deoxyribonucleic acid measurements of cells from the tumor spheroids and from biopsy material obtained directly from the operation revealed the same ploidy and the same amount of proliferating cells in the spheroids as in the original tumor. Fluorescence microscopy using bromodeoxyuridine (BUdR) incorporation and anti-BUdR monoclonal antibody confirmed the proliferative potential of tumor cells in the spheroids. Diameter measurements showed that the size of the spheroids from two of the tumors increased over time while in three other cases it decreased. Spheroids from the remaining two tumors showed no change in size, even after 80 days in culture. These growth data and the relatively high number of proliferating cells, as measured by flow cytometry, indicate that the degree of cell proliferation and cell loss from the spheroids are closely linked, as is the case for tumors in vivo. The culture system presented provides a valuable alternative to propagation of human tumors in animals.

Cellular effects of oncolytic viral therapy on the glioblastoma microenvironment

“…18,19 Using a different approach, glioblastoma (GBM) biopsy material obtained at surgery may be cultured in the form of multicellular spheroids, where the essential phenotypic characteristics of the tumor tissue are retained. 20 Direct transplantation of such 'organotypic' spheroids into rodent brains forms lesions that eventually recapitulate all the histopathological hallmarks of GBM in situ, such as extensive single-cell infiltration, angiogenesis the occurrence of necroses with palisading tumor cells, endothelial proliferations and dilated, thrombotic vessels. 18,21 We have recently used this model to evaluate gene transfer to human GBM cells 22,23 as well as to study the biological aspects of tumor cell invasion.…”

HSV Recombinant Vectors for Gene Therapy

“…They consist entirely of tumour cells and therefore fall to represent the native 3-D architecture and heterogeneity of solid tumours in vivo . As an example, glioblastoma (GBM) biopsy material obtained at surgery, has been cultured in the form of multicellular spheroids, where the essential phenotypic characteristics of the tumour tissue are retained [351]. Direct transplantation of such “organotypic” spheroids into rodent brains forms lesions that eventually recapitulate all the histopathological infiltration, angiogenesis, endothelial proliferations and dilated, thrombotic vessels [352].…”

Intratumoral immunotoxin treatment of human malignant brain tumors in immunodeficient animals

“…Brain tumor tissue from patients undergoing craniotomy was cut into small fragments and cultured according to the method described by Bjerkvig et al 10 Briefly, the tissue samples were dis- 847 sected out at surgery and transferred to tubes containing tissue culture medium. Macroscopically representative tissue was selected for culture and in parallel for histopathologic diagnosis.…”