Characterization of serum-free ex vivo-expanded hematopoietic stem cells derived from human umbilical cord blood CD133(+) cells.
The development of ex vivo expansion of hematopoietic stem cells (HSCs) is a promising approach to restore the required bone marrow function of patients with hematological disorders. Previously, we have reported the development of an optimized serum-free and cytokines-limited defined medium using statistic methodology for umbilical cord blood-derived HSC expansion. The aim of this study was to analyze further the characteristics and functions of cells in vitro and in vivo when cultured in this defined medium. After a 7-day batch culture, the average absolute fold expansions for CD133(+) cells, CD34(+)CD133(+) cells, CD34(+)CD38() cells, CD133(+)CD38(-) cells, CD34(+)CXCR4(+) cells, CD133(+)CXCR4(+) cells, and long-term culture-initiating cells were 21-, 20-, 723-, 618-, 160-, 384-, and 8-fold, respectively. The high enrichment of CD38(-) cells and CXCR4(+) cells of the CD34(+) subpopulation provided a very early uncommitted HSC proliferation and homing ability. Furthermore, the expanded cells showed a high level of telomerase activity to maintain their telomere length and repopulated the lethally irradiated NOD/SCID mice in vivo. These results indicated that the cytokines limited expanded cells from CD133(+) cells could substantially support simultaneous expansion of various stem/progenitor cells and engraft with the expanded cells from a low number of HSCs initially.
Yao
Chao-Ling
CL
Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan.
Feng
Yin-Hsun
YH
Lin
Xi-Zhang
XZ
Chu
I-Ming
IM
Hsieh
Tzu-Bou
TB
Hwang
Shiaw-Min
SM
D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000097:blood*; D018952:Antigens, CD34 / Q000097:blood; D001853:Bone Marrow / Q000648:ultrastructure*; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D005312:Fetal Blood / Q000502:physiology*; D006023:Glycoproteins / Q000097:blood*; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D010455:Peptides / Q000097:blood*; D013234:Stem Cells; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000737:chemistry
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D016207:Cytokines; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D019098:Telomerase