Enrichment of colony forming cells by methotrexate and isokinetic density gradient centrifugation.
Purification of target cells responsive to granulopoietic regulators would be of importance in understanding their mechanism of action. Previous studies have shown that mouse marrow could be enriched 3- to 7-fold with respect to agar colony forming cells (CFU-C) by appropriately scheduled doses of Methotrexate. To further increase this enrichment, such marrow has been subjected to centrifugation through an isokinetic gradient of Ficoll in culture medium. Under optimal conditions (17.5 min at 77.5 X g) the combination of chemotherapy and velocity sedimentation yielded a 17-fold enrichment of CFU-C's.
D000818:Animals; D001853:Bone Marrow / Q000187:drug effects*; D001854:Bone Marrow Cells; D002455:Cell Division / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D002999:Clone Cells / Q000187:drug effects; D005260:Female; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D007279:Injections, Subcutaneous; D008297:Male; D008727:Methotrexate / Q000008:administration & dosage / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D013268:Stimulation, Chemical
D008727:Methotrexate