Murine long-term repopulating ability is compromised by ex vivo culture in serum-free medium despite preservation of committed progenitors.
Hematopoietic progenitor cells can be maintained and expanded ex vivo in standard or serum-free culture medium supplemented with a variety of stimulatory cytokines. The use of serum-free medium allows specification of reproducible and precise growth conditions optimal for various applications and is more acceptable from a safety and regulatory point of view. Human and murine committed progenitor cells have been shown to be equivalently ro better supported by serum-free culture conditions in the presence of multicytokine combinations, but there is little information on the effects of such culture conditions on repopulating stem cells. We used a murine competitive repopulation model to assess the effect of serum-free versus serum-containing ex vivo culture on long-term reconstituting cells. Despite equivalent numbers of committed CFU-C and day 12 CFU-S present after 4 days of culture of murine marrow in serum-free or serum-containing conditions in the presence of IL-3, IL-6, and SCF, long-term reconstituting activity was significantly impaired by serum-free culture. These findings may have important implications for transplantation and gene therapy applications.
Sekhar
M
M
Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Yu
J M
JM
Soma
T
T
Dunbar
C E
CE
D000818:Animals; D001793:Blood Preservation; D001797:Blood Protein Electrophoresis; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured / Q000502:physiology* / Q000637:transplantation; D016895:Culture Media, Serum-Free; D018380:Hematopoietic Stem Cell Transplantation; D006454:Hemoglobins / Q000032:analysis; D007377:Interleukin-3 / Q000502:physiology; D015850:Interleukin-6 / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D019089:Stem Cell Factor / Q000502:physiology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*
D016895:Culture Media, Serum-Free; D006454:Hemoglobins; D007377:Interleukin-3; D015850:Interleukin-6; D019089:Stem Cell Factor