1994 Volume 40 Issue 5 Pages 694-701
Human umbilical cord blood (HUC) contains as many hematopoietic stem/progenitor cells as bone marrow. However, this number may not be sufficient for allogenic transplantation in adults. In this study, mononuclear cells from umbilical cord blood were loaded into flasks of the AIS Micro CELLector™ to capture CD34+ cells. CD34+ cells were enriched 60 times by the treatment and the recoveries of CD34+ cells and colony forming cells was 37.3±19.3% and 14.5±11.2% (n=14, mean±SD), respectively. The enriched CD34+ cells were then cultured in vitro for 7 to 14 days in the presence of IL-3, IL-6 and stem cell factor. The total number of cells and the number of granulocyte-macrophage colony-forming cells increased 80.8±51.2 and 75.0±53.6 fold, respectively in 14 days. The number of CD34+ cells was retained up to 7 days and increased 3-fold after 14 days. Surface marker analysis showed that the percentage of CD34+ CD33- cells decreased from 65.5% to 4.0% in 3 days and that of CD34- CD33+ cells increased from 3.8% to 86.2% in 7 days. The percentage of CD34+ CD38- cells decreased from 10.6% to 3.8% in 3 days and that of CD34- CD38+ cells increased from 10.9 to 76.2% in 7 days. These results indicate that the cultured progenitor cells differentiated as the culture period was extended.
Thus it was shown that hematopoietic progenitor cells enriched from HUC using the Micro CELLector system can be efficiently expanded in vitro.
