A two-step procedure for obtaining 80-fold enriched suspensions of murine pluripotent hemopoietic stem cells.
A method to enrich for murine pluripotent hemopoietic stem cells is described. It consists of two steps. Cells with a density of less than 1.072 g/cm3 at 4 degrees C are first selected from mouse bone marrow by a single-step bovine serum albumin density separation. These low-density cells are then labeled with fluoresceinated wheat germ lectin and sorted by a light-activated cell sorter on the basis of differences in light scatter and fluorescence intensities. Numbers of pluripotent stem cells were determined by the spleen colony assaY (CFU-S). The concentration of CFU-S was 80 times higher in the sorted cell suspension than in the unfractionated control.
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D037102:Lectins; D051379:Mice; D012710:Serum Albumin, Bovine; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins
D005452:Fluoresceins; D037102:Lectins; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate