Immunoselection of oligodendrocytes by magnetic beads. I. Determination of antibody coupling parameters and cell binding conditions.
Oligodendrocytes from early postnatal mouse cerebellum were isolated using polyacrylamide-coated magnetic beads carrying monoclonal antibody to 04 cell surface antigen. Oligodendrocytes were enriched to a purity of 91 +/- 4% starting from a mixed cell population containing approximately 1.5% antigen-positive oligodendrocytes. Viability of 04 antigen-positive oligodendrocytes was approximately 90% as judged by exclusion of trypan blue. Oligodendrocytes were recovered after detachment from the beads with a yield of 19 +/- 6% and after collection by centrifugation onto glass coverslips with yields of approximately 6% of all 04 antigen-positive cells. The final cell yield of oligodendrocytes is approximately 8 x 10(5) cells/gram fresh cerebellar tissue.
D000178:Acrylamides; D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology*; D002846:Chromatography, Affinity / Q000379:methods; D005455:Fluorescent Antibody Technique; D006454:Hemoglobins; D007136:Immunoglobulins; D008280:Magnetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009457:Neuroglia / Q000166:cytology*; D009836:Oligodendroglia / Q000166:cytology*
D000178:Acrylamides; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006454:Hemoglobins; D007136:Immunoglobulins