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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 Publisher Pubmed



Mobarra N1, 2, 3 ; Soleimani M4 ; Ghayourmobarhan M5 ; Safarpour S2 ; Ferns GA6 ; Pakzad R7, 8, 9 ; Pasalar P10
Authors
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Authors Affiliations
  1. 1. Department of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran
  2. 2. Department of Clinical Biochemistry, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
  3. 3. Stem Cell Research Center, Golestan University of Medical Sciences, Gorgan, Iran
  4. 4. Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
  5. 5. Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
  6. 6. Brighton and Sussex Medical School, Division of Medical Education, Brighton, United Kingdom
  7. 7. Noor Research Center for Ophthalmic Epidemiology, Noor Eye Hospital, Tehran, Iran
  8. 8. Department Biostatistics, Faculty of Health, Ilam University of Medical Sciences, Ilam, Iran
  9. 9. Department of Epidemiology, Faculty of Health, Ilam University of Medical Sciences, Ilam, Iran
  10. 10. Metabolic Disorder Research Center, Endocrinology and Metabolism, Molecular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran

Source: Journal of Cellular Physiology Published:2019


Abstract

A suitable alternative strategy for liver transplantation is the use of nanofibrous scaffolds together with stem cells. In this study, a random hybrid of poly-l-lactic acid (PLLA) and poly(ε-caprolactone) (PCL) was used as a three-dimensional (3D) culture for differentiation of hepatocyte-like cells and compared with routine culture (two-dimensional [2D]). The expression of the endodermal marker, forkhead box A2 (FOXA2), was assessed on Day 3 and the hepatic markers; albumin (ALB), α-1 antitrypsin (AAT), and cytokeratin-18 (CK-18) were evaluated on Day 18 using quantitative polymerase chain reaction qPCR. As well as, ALB, α-fetoprotein (AFP), and low-density lipoprotein (LDL) uptake were evaluated using immunocytochemistry; moreover, periodic acid-Schiff and Oil Red were done by cell staining. In addition, AFP and urea production were evaluated by chemiluminescence and colorimetric assays. Light and scanning electron microscopy (SEM) showed changes in the cells in 2D and 3D models. The gene expression of hepatic markers was significantly higher in the 3D cultures. In addition, immunocytochemistry and cell staining showed that ALB, AFP, LDL-uptake, periodic acid-Schiff, and Oil Red were expressed in both cells derived on 2D and 3D. Furthermore, the evaluation of AFP and urea secretion was significantly different between 2D and 3D strategies. These findings suggest that functionally cells cultured on a PLLA/PCL scaffold may be suitable for cell therapy and regenerative medicine. © 2018 Wiley Periodicals, Inc.
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