CRYOPRESERVATION OF ORGANOTYPIC MULTICELLULAR SPHEROIDS FROM HUMAN GLIOMAS

Fresh human glioma tissue can be cultured on agarose to form organotyp ic multicellular spheroids (OMS). The major advantage of OMS is the pr eservation of the cellular heterogeneity and the tumour architecture, which is lost in conventional monolayer cultures. The present study wa s undertaken to assess the possibilities of storing frozen OMS from se ven gliomas which were frozen to determine the viability after thawing . OMS were frozen slowly to -196 degrees C using a programmable freezi ng machine in culture medium containing 45% serum and 10% of the cryop reservative agent dimethyl sulphoxide (DMSO). After 2 weeks storage at -196 degrees C, quick thawing, and culturing for another week, it app eared that the frozen-thawed OMS were viable and retained their histol ogical characteristics. In addition, it is demonstrated that the cellu lar constituents of the OMS resumed metabolic and proliferative activi ties. It is concluded that it is possible to establish frozen stocks o f viable glioma OMS. This will enable extensive studies on OMS, such a s investigation of the biological behaviour of gliomas by using OMS ob tained from primary and corresponding recurrent gliomas. In addition, cryopreservation of OMS makes it possible to correlate the results of in vitro tests on OMS with the patients' responses to similar therapeu tic approaches.