Hematopoietic developmental potential is variable in pluripotent stem cell lines

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TESAŘOVÁ Lenka ŠALINGOVÁ Barbara ŠIMARA Pavel KRONTORÁD KOUTNÁ Irena

Rok publikování 2015
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

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Popis The clinical application of hematopoietic precursors generated from human pluripotent stem cells (hPSCs) is still limited by low efficient differentiation protocols and functional defects in the derived cells. Moreover, the potential of various hPSCs to differentiate into blood cells has not been examined properly. In this study, protocols for hematopoietic differentiation were applied to available human embryonic stem cell (hESC) and human induced pluripotent stem cell (hiPSC) lines to determine their hematopoietic developmental potential. These protocols included different methods for hPSC cultivation and embryoid bodies (EBs) formation and composition of differentiation media. The efficiency of hematopoietic differentiation was found to be variable among hPSC lines. No protocol optimization enabled generation of CD34+ hematopoietic precursors from available hESC lines CCTL-12 and CCTL-14. On the contrary, these precursors were detected during hiPSC differentiation in basic media supplemented with three cytokines. The yield of precursors was variable, ranging from 0.8 to 10.3 percent of CD34+ cells and from 1.0 to 10.6 percent of CD43+ cells at day seven of differentiation. This yield was further increased when hiPSCs were differentiated for ten days in media with richer cytokine supplement. Nevertheless the variability among hiPSC lines was retained, 6.4 to 16.3 percent of cells were CD34+ and 7.8 to 20.0 percent of cells were CD43+235+ precursors of primitive hematopoiesis, while CD45+ precursors of definitive hematopoiesis comprised only from 0.4 to 4.7 percent. These results indicate that hematopoietic developmental potential of individual hPSC lines is characterised by significant variability that has to be overcome before blood cells derived from hPSC could be used for clinical applications.
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