Improvement of hepatogenic differentiation of iPS cells on an aligned polyethersulfone compared to random nanofibers

Abstract

The application of stem cells holds great promises in cell and tissue transplants. This study was designed to compare the hepatogenic differentiation of iPSCs on aligned PES/COL versus random. Aligned and random PES/COL nanofibrus scaffolds were fabricated by electrospining and their surface modified through plasma treatment and collagen coating. The scaffolds were characterized using scanning electron microscopy (SEM) and ATR-FTIR. Morphology and biochemical activities of the differentiated hepatocyte-like cells (HLCs) were examined after 5 and 20 days of differentiation. Real-Time RT-PCR and ICC showed no significant difference in the mRNA and protein levels of two important definitive endoderm specific markers, including Sox17 and Foxa2 between two scaffolds. However, Real-Time RT-PCR analysis indicated an increase in the expression of Cyp7A1 gene over the period of the differentiation procedure on the aligned nanofibers but there was no difference in other genes such as Albumin and CK19. Moreover, comparison of hepatogenic differentiation evaluated by Albumin production in conditioned media of HLCs differentiated on aligned PES/COL, showed increase expression of these markers after 20 days compared to that of the random nanofibers. Taken together, the results of this study may indicate that aligned PES/COL nanofibrous scaffolds can improve terminal differentiation of HLCs from iPSCs.

Hybrid poly‐l‐lactic acid/poly(ε‐caprolactone) nanofibrous scaffold can improve biochemical and molecular markers of human induced pluripotent stem cell‐derived hepatocyte‐like cells

“…We found that (a) PLLA/PCL polymer supports the attachment, proliferation, and differentiation of hiPSCs; (b) the successful generation of hepatic‐like cells from hiPSCs on the hybrid scaffold; and (c) the newly derived cells expressed higher levels of liver markers than cells differentiated on gelatin‐coated plates and single nanofibrous scaffolds. Our results are consistent with those of Mahmoodinia Maymand et al (), in using SEM, we found that the formed cells aggregated, adhered, and penetrated to the scaffold. Our SEM results showed a highly porous and nonwoven architecture that mimics ECM structure, likewise Kinasiewicz et al ().…”

“…Preliminary studies have used various scaffolds such as PES (Hashemi et al, ; Kazemnejad et al, ), PLLA (Feng et al, ; Ghaedi et al, ), and PCL/collagen/PES (Mahmoodinia Maymand et al, ) for hepatocyte differentiation. In the current study, our results demonstrated that hepatic markers are markedly expressed better in hybrid scaffolds than single nanofibrous scaffolds (Mahmoodinia Maymand et al, ). l ‐Form of poly‐lactic acid is a polyester approved by Food and Drug Administration and l ‐lactic acid is the product of this scaffolds degradation, which is harmless for the body (Islami, Mortazavi, Soleimani, & Nadri, ).…”

Decellularized Wharton's jelly extracellular matrix as a promising scaffold for promoting hepatic differentiation of human induced pluripotent stem cells

“…14,26,43 To investigate the effect of DWJS on hiPSCs during hepatic differentiation, hiPSCs were cultured on DWJS or in TCPs with same hepatic induction medium after which cell morphology and expression of specific hepatic markers were examined. 14,26,43 To investigate the effect of DWJS on hiPSCs during hepatic differentiation, hiPSCs were cultured on DWJS or in TCPs with same hepatic induction medium after which cell morphology and expression of specific hepatic markers were examined.…”

“…22,23 Using xenogeneic primary hepatocyte and immortalized hepatocellular carcinoma cell lines are also associated with some problems, including immunogenicity, pathogenicity, and tumorgenesis. 23,[25][26][27] Recently, decellularized scaffold of heart and lung were repopulated with human iPSC-derived organ specific cells. Due to the unlimited selfrenewing capacity and less immunogenicity of iPSCs, they became a preferred choice for hepatocyte generation.…”

Generation of insulin‐producing cells from human adipose‐derived mesenchymal stem cells on PVA scaffold by optimized differentiation protocol

“…Up to recent years, the IPCs differentiation of hADSCs has mostly been performed in 2D culture and fully functional and mature islet‐like clusters maintaining their features in the condition of 3D culture have not been developed. In addition, natural and polymeric synthetic scaffolds provide a suitable substrate and surface area for cell seeding and attachment as well as developing and improving stem cell proliferation and differentiation capability (Amer, Mahoney, & Bryant, ; Mahmoodinia et al, ). The combination of hADSCs with nanofibrous scaffolds has many useful effects including the slow diffusion of growth factors and metabolites, as well as the fabrication of pancreas architecture for pancreas tissue engineering purposes in the future (Javad et al, ).…”