IMMUNOMAGNETIC ISOLATION OF HUMAN DEVELOPING MOTOR-NEURONS

HUMAN motor neuron (MN) isolation provides a critical tool to study ne urophysiological properties and the effects of molecules of clinical r elevance on isolated neurons. We developed an immunomagnetic separatio n technique based on specific MN antigen recognition for nerve growth factor receptor (p(75-NGFR)). We cultured an average of 250 000 cells from the anterior horns of a single cord (four specimens at postconcep tion Weeks 6.0, 7.2, 8.0, and 8.3). At day 7 in vitro (DIV), choline a cetyltransferase (ChAT) and/or p(75-NGFR)-expressing cells (MNs) repre sented 72 +/- 2% of the total growing cells. MNs survived for at least 4 weeks in biochemically defined medium. The immunomagnetic separatio n method has been demonstrated to be effective, reproducible, and quan titative for separation of MNs. (C) 1998 Rapid Science Ltd.