Immortalized neuroectodermal precursor cell lines were generated from mouse brain by the SV40 large T antigen expressed under the control of the LAP (lac activating protein) mammalian regulatory system. The LAP system permits the reversible expression of T antigen as a function of the exogenous inducer, isopropyl‐β‐D‐thiogalactopyranoside. Immortalized cells can be stably maintained in an undifferentiated state in monolayer cultures. Cell lines expressed the early neurofilament‐like protein nestin, but not markers characteristic for mature cells such as the neurofilament light protein and glial fibrillary acidic protein. Downregulating the LAP‐controlled T antigen with isopropyl‐β‐D‐thiogalactopyranoside was not sufficient to induce differentiation. However, when cells formed three‐dimensional aggregates, differentiation to a neuronal phenotype occurred, indicating that cell‐cell interaction plays an important role in their differentiation. Cells in aggregates did not proliferate, even in the presence of T antigen, suggesting that an aggregation‐induced signal to cease growth was dominant over the growth signal of T antigen. Further morphological differentiation was induced by basic fibroblast growth factor. These immortalized cells should facilitate molecular and cellular studies concerned with the mechanism of commitment, fate determination, and mitotic arrest of neuronal precursor cells in the developing mammalian CNS. © 1995 Wiley‐Liss, Inc.
All Science Journal Classification (ASJC) codes
- Cellular and Molecular Neuroscience
- T antigen