Long-term, EGF-stimulated cultures of attached GFAP-positive cells derivedfrom the embryonic mouse lateral ganglionic eminence: In vitro and transplantation studies

Long-term attached cultures, prepared from mouse embryonic days 15-17 later al ganglionic eminence, were grown in a medium including epidermal growth f actor and serum, and the survival, differentiation, and migration of cells from either early or late passages were analyzed following transplantation. The cultured cells had the morphology of type I astroglial cells, with the vast majority of the cells immunoreactive for glial fibrillary acidic prot ein (around 90%), the intermediate filament marker nestin, and also the mou se-specific neural markers M2 and M6. The cultures were kept over 25 passag es (7 months). During the first 8 passages, the growth rate gradually decli ned, but it increased again after passage 9 and thereafter stabilized at va lues similar to those observed during the initial culture period. After pas sages 4-6 and 18, cell suspensions were implanted cross-species into the in tact or lesioned striatum of adult (passages 4-5 only) or intact striatum o f neonatal rats (passages 4-6 or 18). Both early and late passage cells for med M2 (and M6)-positive transplants. In the neonatal recipients, widesprea d migration was seen from the, needle tract throughout most of the striatum , along the internal capsule, and into the globus pallidus. In the adult st riatum, the cells remained mostly around the injection tract, or within 0.4 -0.6 mm from the graft core. These long-term attached cultures are interest ing to compare to nonattached neurosphere cultures, and might also offer a means of propagating relatively pure populations of astroglia-like cells fo r basic transplantation studies or for use in experimental trials with ex v ivo gene transfer. (C) 2000 Academic Press.