Establishment of a permanent rat brain-derived glial cell line as a source of purified oligodendrocyte-type 2 astrocyte lineage cell populations.
A permanent glial cell line (L3) has been established from mixed glial cultures obtained from neonatal rat forebrain by repetitive passaging and selection of the process-bearing cells growing on top of a flat cell monolayer. Continuous propagation of the process-bearing cells was supported by the flat cells, of presumed astroglial origin, which were present in negligible amounts following each passage but then grew and formed a basal, feeder layer. Throughout a culture period of over 2 years, the L3 cells have maintained a stable morphological and antigenic phenotype. In serum-containing culture medium, most of the process-bearing cells expressed at the same time features of immature oligodendrocytes (O4 positivity) and of astrocytes [glial fibrillary acidic protein (GFAP) positivity]. A smaller proportion of them was labeled by the monoclonal antibody LB1. LB1+ or O4+ cells were rarely GFAP-, and GFAP+ cells were rarely LB1- or O4-. GalC+ oligodendrocytes were seen only occasionally, but the proportion of these cells increased up to 30% upon culturing in chemically defined medium containing 0.5% fetal calf serum. The L3 process-bearing cells accumulated the neurotransmitter gamma-aminobutyric acid (GABA), expressed the proteoglycan chondroitin sulfate, and responded to the mitogenic action of platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF). All these properties are characteristic of cells belonging to the O-2A (oligodendrocyte-type 2 astrocyte) cell lineage. The L3 flat cells were largely negative for the glial markers tested, but resembled type 1 astrocytes in their ability to support the growth of O-2A lineage cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Aloisi
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F
Laboratory of Organ and System Pathophysiology, Istituto Superiore di Sanità, Rome, Italy.
Sun
D
D
Levi
G
G
Wekerle
H
H
D000818:Animals; D000831:Animals, Newborn; D000941:Antigens / Q000032:analysis; D001253:Astrocytes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D001921:Brain / Q000166:cytology*; D002455:Cell Division; D002460:Cell Line; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006133:Growth Substances / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D000941:Antigens; D005904:Glial Fibrillary Acidic Protein; D006133:Growth Substances; D005680:gamma-Aminobutyric Acid