Isolation in a single step of a highly enriched murine hematopoietic stem cell population with competitive long-term repopulating ability

摘要: A simple procedure is described for the quantitation and enrichment of murine hematopoietic cells with capacity long-term repopulation lymphoid myeloid tissues in lethally irradiated mice. To ensure detection most primitive marrow this potential, we used a competitive assay which female recipients were injected male test 1 to 2 x 10(5) compromised normal short-term repopulating ability, but whose ability had been reduced by serial transplantation. Primitive purified flow cytometry sorting based on their forward orthogonal light-scattering properties, Thy-1 H-2K antigen expression. Enrichment profiles marrow, mice 5-fluorouracil (5-FU) four days previously, established each these parameters using an vitro high proliferative pluripotent colony-forming cells. When all gated simultaneously, clonogenic enriched 100-fold. Both day 9 12 CFU-S copurified; however, purity (23%) (75-fold) sorted population was greater 5-FU-treated Five hundred 5-FU consistently repopulated recipient (greater thanmore » 50% male, 3 months post-transplant) when co-injected cells, approximately 100 sufficient achieve same result under conditions. This relatively purification strategy should facilitate further analysis heterogeneity regulation stem that maintain hematopoiesis vivo.« less