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dc.contributor.authorPranke, Patricia Helena Lucaspt_BR
dc.contributor.authorHendrikx, Janpt_BR
dc.contributor.authorDebnath, Gargipt_BR
dc.contributor.authorAlespeite, Gabrielpt_BR
dc.contributor.authorRubinstein, Pablopt_BR
dc.contributor.authorNardi, Nance Beyerpt_BR
dc.contributor.authorVisser, Jan W. M.pt_BR
dc.date.accessioned2010-04-24T04:15:39Zpt_BR
dc.date.issued2005pt_BR
dc.identifier.issn0100-879Xpt_BR
dc.identifier.urihttp://hdl.handle.net/10183/21187pt_BR
dc.description.abstractIdentification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO), FLT-3 ligand (FL) and kit ligand (KL; or stem cell factor) in different combinations: TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9% CD34+ cells, 2.6 ± 2.1% of which were CD38- negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25%. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29% of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype.en
dc.format.mimetypeapplication/pdfpt_BR
dc.language.isoengpt_BR
dc.relation.ispartofBrazilian journal of medical and biological research = Revista brasileira de pesquisas médicas e biológicas. Ribeirão Preto, SP. Vol. 38, no. 12 (Dec. 2005), p. 1775-1789pt_BR
dc.rightsOpen Accessen
dc.subjectHematopoietic stem cellsen
dc.subjectCélulas-tronco hematopoéticaspt_BR
dc.subjectHematologiapt_BR
dc.subjectCD34+CD38- cellsen
dc.subjectHuman umbilical cord blooden
dc.subjectEx vivo expansionen
dc.subjectAdhesion moleculesen
dc.titleImmunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culturept_BR
dc.typeArtigo de periódicopt_BR
dc.identifier.nrb000515797pt_BR
dc.type.originNacionalpt_BR


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