Abstract
Cell therapy and tissue engineering as promising candidates for the liver transplantation dilemma are of special interest. Induced pluripotent stem cells (iPSCs) are one of the best sources in this field, but their differentiation methods to hepatocytes have remained challenging. We transduced human iPSCs (hiPSCs) with miR‐122 and off‐let‐7f (hiPSCsmiR‐122 + off‐let‐7f) to evaluate how they can differentiate hiPSCs to hepatocyte‐like cells (HLCs) without any extrinsic growth factor. Additionally, we studied the effect of Poly ɛ‐caprolactone‐gelatin‐hyaluronic acid (PCL‐Gel‐HA) nanofibrous scaffold as an extracellular matrix (ECM) simulator on differentiation improvement. Definitive endoderm markers (FOXA2 and SOX17), as well as hepatic markers (AFP, Albumin, CK18, HNF4α) expression, were significantly higher in hiPSCsmiR‐122 + off‐let‐7f derived HLCs (hiPSCs‐HLCs) compared to the control group (miR‐scramble transduced hiPSCs: hiPSCsscramble). hiPSCs‐HLCs indicated hepatocyte morphological characteristics and positive immunostaining for AFP, Albumin and HNF4α. Albumin and urea secretion were significantly higher in hiPSCs‐HLCs than hiPSCsscramble. Comparing these markers in the PCL‐Gel‐HA group with the tissue culture plate (TCP) group revealed that PCL‐Gel‐HA could improve differentiation towards HLCs significantly. Regarding our results, these microRNAs can be used to differentiate hiPSCs to the functional hepatocytes for disease modelling, drug screening and cell‐based therapy in future studies.
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