Isolation of murine pluripotent hemopoietic stem cells in the Go phase.
A method to purify pluripotent hemopoietic stem cells in the Go phase from mouse bone marrow was established. Bone marrow cells from 5-fluorouracil (5-FU)-treated mice were fractionated by Percoll density gradient. The cells with density between 1.063 and 1.075 were further separated into wheat germ agglutinin (WGA)-positive and -negative cells using fluorescent-activated cell sorter (FACS) after staining with fluorescein isothiocyanate-conjugated WGA (FITC-WGA). An assay for spleen colony-forming units (CFU-S) revealed that the WGA-positive cells (1 X 10(6)) produced 1380 CFU-S (about 150 times of the number in the original bone marrow cells) on day 12 (but no CFU-S on day 8), whereas the WGA-negative cells produced no CFU-S. Thus, the stem cells in the Go phase are found to be enriched 150 times in 5-FU-treated WGA-positive cells.
D000818:Animals; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D003114:Colony-Forming Units Assay; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D005452:Fluoresceins; D005472:Fluorouracil / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007399:Interphase; D051379:Mice; D008809:Mice, Inbred C3H; D008854:Microscopy, Electron; D014909:Wheat Germ Agglutinins
D005452:Fluoresceins; D014909:Wheat Germ Agglutinins; C047939:fluorescein isothiocyanate-wheat germ agglutinin; D016650:Fluorescein-5-isothiocyanate; D005472:Fluorouracil