Roslan, Fatin Fazrina
(2024)
Optimisation Of Induced Pluripotent Stem Cells Derivation From Human Cord Blood And Its Application For Cardiac Regeneration In Cryoinjured Rat Model.
PhD thesis, Universiti Sains Malaysia.
Abstract
Cardiomyocyte therapy was considered the only option to repopulate myocardial loss, but challenges existed in achieving effective replenishment of functional cardiomyocytes that could translate into clinical benefits. This study aimed to examine the use and function of pre-epicardial cells in cardiomyocyte therapy in vivo. Cryopreserved cord blood was used to generate human-induced pluripotent stem cells (hiPSCs) to harness the ‘youthfulness’ of foetal cells. The generated hiPSCs were also used to produce cardiomyocytes (CMs) and pre-epicardial cells (PECs) with high differentiation efficiency using established protocols. To confirm the effects of PECs on cardiomyocytes, both differentiated CMs and PECs were co-cultured for 6 days, and the proliferation and role of Hippo signaling were assessed. PECs (1x10⁶ cells/100 μl) were also combined with CMs (1x10⁶ cells/100 μl) to treat cryoinjured rat hearts after 1 month. Cardiac functions were assessed using Millar Pressure Volume Loop cardiac hemodynamics analysis. Differences between groups were considered significant when p < 0.05 using ANOVA. Frozen CD34 cells were successfully reprogrammed into hiPSCs with consistent expression of pluripotent markers. The generated hiPSCs demonstrated multilineage differentiation ability, forming cardiomyocytes, dopamine neurons, and early endodermal cells.
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