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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
journal contribution
posted on 2023-06-10, 03:44 authored by Naser Mobarra, Sara Raji, Sara Najafi, Farzaneh Kamelan Kafi, Gordon FernsGordon Ferns, Reza PakzadAn 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.
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- Published
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- Published version
Journal
Oxidative Medicine and Cellular LongevityISSN
1942-0900Publisher
Hindawi LimitedExternal DOI
Volume
2021Page range
1-15Article number
a4229721Event location
United StatesDepartment affiliated with
- Division of Medical Education Publications
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- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2022-05-30First Open Access (FOA) Date
2022-05-30First Compliant Deposit (FCD) Date
2022-05-30Usage metrics
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