조혈간세포의 분리 및 체외증식

초록

Umbilical cord blood is increasingly being used in the setting of allogeneic marrow transplantation. We review the characteristics of hematopoietic stem cells and evaluate the potential of ex vivo expansion from different stem cell sources. After Ficoll-Paque density gradient method and then positive selection of CD34+ cells using magnetic cell sorting and separation method by using Miltenyi MiniMACS column, the CD34+cells from bone marrow (BM), umbilical cord blood (CB), and mobilized peripheral blood (PB) were cultured for BFU-E, CFU-GM and CFU-MK at day 0, day 4, day 7, and day 14 under the combination of growth factors, with cell counts. CB-derived CD34+ cells had significantly higher total cell proliferation than either BM or PB at day 7 and day 14. The colony count of BFU-E was in general more plentiful in CB than in BM and PB at day 4, day 7 and day 14, among which the difference was the most distinct at day 7 culture. Also, CB CD34+ cells produced more CFU-Mk colonies than did BM or PB at day 4 and day 7. Ex vivo expansion of CB cells may be most promising in producing total cellular expansion, CFU-Mk and BFU-E compared with BM and PB, especially at day 7, because the former was the most productive hematopoietic source on a per volume basis.