Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Hypoxia-Induced miR-210 Overexpression Promotes the Differentiation of Human-Induced Pluripotent Stem Cells to Hepatocyte-Like Cells on Random Nanofiber Poly-L-Lactic Acid/Poly (ϵ -Caprolactone) Scaffolds

Fri Aug 19 04:13:47 2022

(2021) Hypoxia-Induced miR-210 Overexpression Promotes the Differentiation of Human-Induced Pluripotent Stem Cells to Hepatocyte-Like Cells on Random Nanofiber Poly-L-Lactic Acid/Poly (ϵ -Caprolactone) Scaffolds. Oxidative Medicine and Cellular Longevity. ISSN 19420900 (ISSN)

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Official URL: https://www.scopus.com/inward/record.uri?eid=2-s2....

Abstract

An alternative treatment to liver transplantation includes the use of differentiated stem cells. Hypoxia has been shown to endow human-induced pluripotent stem cells (hiPSCs) with enhanced hepatic differentiation. We have investigated a new strategy for hepatocyte differentiation from hiPSCs using a three-step differentiation protocol with lentiviral overexpression of hypoxia-microRNA-210 of cells grown on a hybrid scaffold. We analyzed the transduction of the miR-210 lentiviral and definitive endoderm and pluripotency gene markers, including SRY-box 17 (SOX17), forkhead box A2 (FOXA2), and octamer-binding transcription factor 4 (OCT-4) by Real-Time PCR and fluorescent microscope. The scanning electron microscopy (SEM) examined the 3D cell morphological changes. Immunocytochemistry staining was used together with assays for aspartate aminotransferase, alanine aminotransferase, and urea secretion to analyze hepatocyte biomarkers and functional markers consisting of α-fetoprotein (AFP), low-density lipoprotein (LDL) uptake, fat accumulation, and glycogen. The flow cytometry analyzed the generation of reactive oxygen species (ROS). Compared to cells transfected with the blank lentiviral vectors as a control, overexpressing miR-210 was at higher levels in hiPSCs. The expression of endodermal genes and glycogen synthesis significantly increased in the differentiated lentiviral miR-210 cells without any differences regarding lipid storage level. Additionally, cells containing miR-210 showed a greater expression of ALB, LDL, AST, ALT, urea, and insignificant lower AFP and ROS levels after 18 days. However, SEM showed no significant differences between cells under the differentiation process and controls. In conclusion, the differentiation of hiPSCs to hepatocyte-like cells under hypoxia miR-210 may be a suitable method for cell therapy and regenerative medicine. © 2021 Naser Mobarra et al.

Item Type: Article
Creators:
CreatorsEmail
Mobarra, N.UNSPECIFIED
Raji, S.UNSPECIFIED
Najafi, S.UNSPECIFIED
Kafi, F. K.UNSPECIFIED
Ferns, G. A.UNSPECIFIED
Pakzad, R.UNSPECIFIED
Keywords: Amino acids Cytology Lactic acid Metabolism Photoelectric microscopes Polymerase chain reaction RNA Scaffolds (biology) Stem cells Transcription Urea Caprolactone Hepatic differentiation Hepatocytes Human-induced pluripotent stem cells Liver transplantation Low density lipoproteins Overexpressions Poly-l-lactic acids Reactive oxygen species Stem-cell Scanning electron microscopy Alanine Boxes Cells Differentiation Genes Markers Medicine
Divisions:
Journal or Publication Title: Oxidative Medicine and Cellular Longevity
Journal Index: Scopus
Volume: 2021
Identification Number: https://doi.org/10.1155/2021/4229721
ISSN: 19420900 (ISSN)
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/3790

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