OLN-93 - A NEW PERMANENT OLIGODENDROGLIA CELL-LINE DERIVED FROM PRIMARY RAT-BRAIN GLIAL CULTURES

We have established a new permanent cell line (OLN-93), derived from s pontaneously transformed cells in primary rat brain glial cultures. In growth medium supplemented with 10% fetal calf serum a doubling time of 16-18 hr was determined. OLN-93 cells in their antigenic properties resemble primary oligodendrocytes in culture. As analyzed by indirect immunofluorescence, the A2B5 surface marker is absent, they express g alactocerebroside and myelin-specific proteins, such as myelin basic p rotein (MBP), myelin-associated glycoprotein (MAG), proteolipidprotein (PLP), and Wolfgram protein (WP), but do not exhibit astrocytic prope rties, such as the expression of vimentin or the glial fibrillary acid ic protein (GFAP). In their morphological features they resemble bipol ar O-2A-progenitor cells and, when grown at low density or on poly-L-l ysine-coated culture dishes under low serum conditions, immature oligo dendrocytes with a more arborized cell morphology. The cellular proces ses contain microfilaments, while N-CAM/D2 immunoreactivity is localiz ed on the cell surface of the somata and processes. Immunoblot analysi s further confirmed the presence of MAG, WP and MBP immunoreactivity, and the absence of vimentin and GFAP. Only a single MBP isoform (simil ar to 14 kDa) was detectable in the cellular extracts. PLP mRNA expres sion was studied by RT-PCR. The two proteo-lipid-specific mRNAs, DM20 and PLP, were present in OLN-93 cell extracts. Comparisons with embryo nic rat cerebral cells in culture and primary oligodendrocytes suggest that OLN-93 cells in their morphological features and their antigenic properties resemble 5- to 10-day-old (postnatal time) cultured rat br ain oligodendrocytes. Thus, the new cell line described in this study should provide a useful model system to investigate the specific mecha nisms regulating the proliferation and differentiation of oligodendroc ytes in vitro, and the molecular interactions with other cells of the nervous system. (C) 1996 Wiley-Liss, Inc.