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作者：Katrien Vanheusdena, Stefanie Van Coppernolleb, Magda De Smedta, Jean Pluma, Bart Vandekerckhovea作者单位：aDepartment of Clinical Chemistry, Microbiology and Immunology, Faculty of Medicine and Health Sciences, Ghent University, Ghent University Hospital, Ghent, Belgium;bDienst voor het Bloed, Rode Kruis-Vlaanderen, Ghent, Belgium
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【摘要】
: }! L( F2 t9 O! G! f: X S Expansion of hematopoietic stem cells could be used clinically to shorten the prolonged aplastic phase after umbilical cord blood (UCB) transplantation. In this report, we investigated rapid severe combined immunodeficient (SCID) repopulating activity (rSRA) 2 weeks after transplantation of CD34 UCB cells cultured with serum on MS5 stromal cells and in serum- and stroma-free cultures. Various subpopulations obtained after culture were studied for rSRA. CD34 expansion cultures resulted in vast expansion of CD45 and CD34 cells. Independent of the culture method, only the CD34 33 38¨C fraction of the cultured cells contained rSRA. Subsequently, we subfractionated the CD34 38¨C fraction using stem cell markers CD45RA and CD90. In vitro differentiation cultures showed CD34 expansion in both CD45RA¨C and CD90 cultures, whereas little increase in CD34 cells was observed in both CD45RA and CD90¨C cultures. By four-color flow cytometry, we could demonstrate that CD34 38¨C45RA¨C and CD34 38¨C90 cell populations were largely overlapping. Both populations were able to reconstitute SCID/nonobese diabetic mice at 2 weeks, indicating that these cells contained rSRA activity. In contrast, CD34 38¨C45RA or CD34 38¨C90¨C cells contributed only marginally to rSRA. Similar results were obtained when cells were injected intrafemorally, suggesting that the lack of reconstitution was not due to homing defects. In conclusion, we show that after in vitro expansion, rSRA is mediated by CD34 38¨C90 45RA¨C cells. All other cell fractions have limited reconstitutive potential, mainly because the cells have lost stem cell activity rather than because of homing defects. These findings can be used clinically to assess the rSRA of cultured stem cells. 8 z# `0 j7 E+ @( s, g
【关键词】 Hematopoiesis Stem cell Stem cell culture Nonobese diabetic/severe combined immunodeficient mice
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发表于 2009-3-5 01:00
发表于 2015-5-23 11:30
