POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS

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ŠIMARA Pavel KRONTORÁD KOUTNÁ Irena TESAŘOVÁ Lenka FARKAŠ Šimon

Rok publikování 2017
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

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Popis Human induced pluripotent stem cells (hiPSCs) are currently considered as the cells which will be widely used in the regeneration of many tissues and organs. One possible utilization is their differentiation into the vascular endothelium. In our laboratory, we have derived hiPSCs from four different cell types. For derivation of hiPSCs we have used as a source cells: neonatal fibroblast, PBMC fraction, HUVECs and HSVECc. On these four hIPSCs lines we tested and evaluated two types of differentiation protocols into endothelial cells. First protocol “A” was aimed on direct differentiation into the lateral mesoderm with subsequent differentiation into the endothelium. In second protocol “B” we have incorporated step of controlled support of differentiation into primitive streak. In case of the first protocol, only up to 31% of cells had desired surface markers (SM) prior to separation. The second protocol yielded up to 48% of cells with desired SM. Quality of differentiated cells in following passages also varied greatly. There was a significant emergence of fibroblast-like cell (FLC) in cell culture by protocol “A”. In such cases, if endothelial cells are not rescued via separation based on SM CD31/CD144 they are overgrown by FLC in matter of days. In passages following initial separation, the cells had 60-80 % of desired SM for the most part, they also needed to be re-separated to maintain these levels of SM. Even though FLC sometimes appeared in cell cultures by protocol “B”, they grew at much slower pace and the rescue separation was as a result much less urgent. Levels of desired SM were also steadily rising even without separation up to 99%. These cells were positive for SM CD31, CD144, CD304 and CD34. Thus differentiated cells showed 100% efficiency on absorption of LDL and tube formation assay. Among the individual hiPSCs lines, we have not found a fundamental difference in the yield between these two protocols.
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