Effect of embryonic donor age and dissection on the DARPP-32 content of cell suspensions used for intrastriatal transplantation.

Abstract

The aim of this study was to determine in vitro the DARPP-32 content of donor cells used for striatal transplantation in vivo. The effect of selective embryonic dissection of the lateral ganglionic eminence (LGE) was compared with the standard dissection of the whole ganglionic eminence (WGE) at each of three embryonic ages (14, 15, and 16 days of gestation) in the rat. The resultant cell suspensions were cultured for up to 7 days and incubated with antibodies against DARPP-32, a marker of striatal medium spiny neurons; beta-tubulin III, a neuronal marker; GFAP, a marker of reactive astrocytes; and Gal-C, a marker of oligodendrocytes. LGE dissection gave rise to more DARPP-32 neurons compared to WGE; but this relationship was only observed in the younger embryos. When older (16 days gestation) embryos are used there is no difference in the yield of DARPP-32 cells obtained from LGE and WGE. LGE dissections were also observed to contain fewer glial cells. There was no beneficial effect of LGE over WGE on survival of striatal neurons in vitro. These results have important implications for the selection and dissection of fetal donor material used in clinical trials of intrastriatal transplantation as a potential treatment for Huntington's disease.