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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 (? -caprolactone) scaffolds

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posted on 2023-06-10, 03:44 authored by Naser Mobarra, Sara Raji, Sara Najafi, Farzaneh Kamelan Kafi, Gordon FernsGordon Ferns, Reza Pakzad
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 a-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.

History

Publication status

  • Published

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  • Published version

Journal

Oxidative Medicine and Cellular Longevity

ISSN

1942-0900

Publisher

Hindawi Limited

Volume

2021

Page range

1-15

Article number

a4229721

Event location

United States

Department affiliated with

  • Division of Medical Education Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2022-05-30

First Open Access (FOA) Date

2022-05-30

First Compliant Deposit (FCD) Date

2022-05-30

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