Long-term expansion of human foetal neural progenitors leads to reduced graft viability in the neonatal rat brain

Abstract

We previously reported that early passage human foetal neural progenitors (hFNPs) survive long-term in the rodent host brain whereas late passage cells disappear at later post-graft survival times. The extent to which this finding is related to changes in the expanded FNPs or in the adult host brain environment was not determined. Here we report the effect of expanding hFNPs for different periods of time in vitro on their ability to survive transplantation into the neonatal rat hippocampus, a generally more permissive environment than the adult rat brain. After 2 and 8 weeks in vitro, transplanted hFNPs formed large grafts, most of which survived well until at least 12 weeks. However, following continued expansion, hFNPs formed smaller grafts, and cells transplanted after 20 weeks expansion produced no surviving grafts, even at early survival times. To determine whether this could be due to a dilution of "true" neural stem cells through more differentiated progeny over time in culture, we derived homogeneous neural stem (NS) cells grown as a monolayer from the 8 week expanded hFNPs. These cells homogeneously expressed the neural stem cell markers sox-2, 3CB2 and nestin and were expanded for 5 months before transplantation into the neonatal rat brain. However, these cells exhibited a similar survival profile to the long-term expanded FNPs. These results indicate that, while the cellular phenotype of neural stem cells may appear to be stable in vitro using standard markers, expansion profoundly influences the ability of such cells to form viable grafts.