Method for separation, morphological and functional characterization of Sertoli's cells from the prepubertal testis: A potential nursing cell system for pancreatic islets

Sertoli's cells (SC) play a major role in the differentiation as well as de velopment of functional competence of several cell populations in the male testis, The biologic properties of these cells span from promotion of cellu lar growth to immunomodulation capacity, although others remain unclear. In order to begin to assess whether SC would be beneficial to pancreatic isle t co-cultures, we have developed an ad hoc method for the reproducible isol ation and morphological and functional characterization of purified SC from the prepubertal animal (rat and pig) testis, We describe an original, fair ly rapid multi-step enzymatic digestion process of the testis which leads t o isolation of 90% pure and viable SC. The non-SC population was purportedl y kept on 10% so as to prevent complete disruption of the testicular inter- cellular cross talk. 90% pure SC have been shown to fully retain morphologi c integrity and viability over several weeks of in vitro culture maintenanc e, where their growth rate has resulted steadily stable. SC, separated from the prepubertal rat testis by this method, have been preliminarily employe d, at particular concentrations, for in vitro SC/islet co-cultures. We have collected preliminary evidence that co-incubation with homologous SC enhan ces insulin output from rat islets as compared with controls. This finding could offer the opportunity to prolong survival and implement functional co mpetence of the islets by providing them with an ancillary nursing cell sys tem. In conclusion, we have set up an original, specifically designed metho d for SC separation with the aim of creating a biologic support for the iso lated islets, (C) 1998, Editrice Kurtis.