PURIFICATION OF CELL-POPULATIONS FROM HUMAN FETAL BRAIN USING FLOW CYTOMETRIC TECHNIQUES

We recently established primary cultures from dissociated second trime ster human fetal brains using a novel spin seeding method and characte rized cellular populations with distinct phenotypes in these cultures. Here, we report that these neural cultures can be dissociated to sing le-cell suspensions, sorted by size using flow cytometry and re-seeded to yield cultures selectively enriched for the neuronal and glial cel l populations. Sorted neurons were highly homogeneous, viable and exte nded processes by one day after re-seeding. These neurons expressed im munoreactivity for neurofilament protein, retained their GABAergic phe notype and were electrically excitable. Re-seeded astrocytes prolifera ted in culture and expressed glial fibrillary acidic protein. We descr ibe the conditions required for the flow cytometric sorting and tissue culture assays as well as the morphological, immunocytochemical and e lectrophysiological characteristics of the sorted neuronal population.