Hematopoietic differentiation of human pluripotent stem cells

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Authors

TESAŘOVÁ Lenka ŠIMARA Pavel ŠALINGOVÁ Barbara KRONTORÁD KOUTNÁ Irena

Year of publication 2014
Type Appeared in Conference without Proceedings
MU Faculty or unit

Faculty of Informatics

Citation
Description Generation of desired cell types from human pluripotent stem cells (hPSCs) promise many applications in regenerative medicine, e.g. in vitro generated hematopoietic progenitors could provide an alternative to hematopoietic stem cells used for transplantations. However, current protocols for hematopoietic differentiation of hPSCs suffer from low efficiency and functional defects in the derived cells. The technology is also limited by an unequal capacity of various hPSC lines to generate hematopoietic progenitors, which could be reasoned by genetic and epigenetic variations of hPSCs. Methodologies for hematopoietic differentiation in feeder-free conditions were applied to available human embryonic stem cell (hESC) and human induced pluripotent stem cell (hiPSC) lines in this study to determine their hematopoietic differentiation potential. At the same time, DNA methylation status of selected genes was evaluated in hPSC lines to find out if this DNA modification is associated with the variability of cell lines and if so called epigenetic memory is maintained in hiPSCs. It was found that different hPSC lines cultured in different conditions develop into embryoid bodies with different efficiency, so for efficient and reproducible hematopoietic differentiation of hPSCs, it is necessary to consider the generation of embryoid bodies itself. Furthermore, available hPSC lines were shown not to generate hematopoietic progenitors in such an extent as H1 and H9 hESC lines used for this purpose in the vast majority of relevant studies, indicating that various hPSCs differ in hematopoietic development potential. In epigenetic analyses, residual promotor DNA methylation of somatic genes was detected in some hiPSC lines depending on the type of reprogramming source cell and the reprogramming technology. In hiPSC and hESC lines variability in DNA methylation level of UTF1 and TSPYL5 was found suggesting these two genes as putative markers linked to the pluripotency state. Our results suggest that for clinical application of blood cells derived from hPSCs, it is necessary to overcome the variability in the hematopoietic developmental potential of individual hPSC lines. The association of DNA methylation status in hPSC lines with their ability to differentiate into blood progenitors will be further studied.
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