In vitro differentiation of E-N-CAM expressing rat neural precursor cells isolated by FACS during prenatal development.
Most fetal rat brain cells expressing the embryonal, highly sialylated form of the cell adhesion molecule N-CAM (E-N-CAM) are precursor cells, as judged from the absence of marker molecules specific for mature neural cell types. However, the detection of E-N-CAM+ cells in frozen sections does not provide information on the lineage-specific differentiation of these cells during development. To investigate their differentiation behaviour in vitro, E-N-CAM+ cells were isolated at different times of brain development by fluorescence-activated cell sorting (FACS), using a monoclonal antibody (Mab RB21-7) which specifically recognizes polysialic acid (PSA) residues on E-N-CAM. Double-immunofluorescence analyses showed that the majority of E-N-CAM+ cells isolated on prenatal days 15 to 18 differentiated into neurons while a small subset of Mab RB21-7 binding cells proved to be astrocytic precursors and/or bipotential. The proportion of E-N-CAM+ astrocytic precursors increased during later development (prenatal day 22) concomitantly with the onset of gliogenesis. While conversion of E-N-CAM to mature forms of N-CAM was never observed in neurons during cultivation, E-N-CAM+ cells of the astrocyte lineage switched to N-CAM soon after the onset of GFAP expression. A lineage-specific transition of E-N-CAM to mature N-CAM expression is, therefore, suggested for these astrocytic progenitor cells during rat brain development.
Blass-Kampmann
S
S
Institute of Cell Biology (Cancer Research), University of Essen Medical School, Germany.
Reinhardt-Maelicke
S
S
Kindler-Röhrborn
A
A
Cleeves
V
V
Rajewsky
M F
MF
D000818:Animals; D000911:Antibodies, Monoclonal; D001253:Astrocytes / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000166:cytology* / Q000196:embryology; D015816:Cell Adhesion Molecules, Neuronal / Q000096:biosynthesis* / Q000276:immunology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D010641:Phenotype; D011233:Precipitin Tests; D051381:Rats; D012794:Sialic Acids / Q000096:biosynthesis; D013234:Stem Cells / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015816:Cell Adhesion Molecules, Neuronal; D012794:Sialic Acids; D013462:Sulfur Radioisotopes; C021319:polysialic acid