Comparison of equine-induced pluripotent stem cell characteristics induced on different cell adhesion substrates.
Abstract: This study evaluated the effects of cell adhesion substrates that lead to the generation of equine-induced pluripotent stem cells (eiPSC) from embryonic skin fibroblasts by lipofection of plasmid vectors expressing five reprogramming factors. The reprogramming efficiency of cells induced on the E8 fragment of laminin-511 (eiPSC-511) was higher than that on Geltrex containing laminin-111 as a major laminin (eiPSC-111), and supplementation with a cocktail of small molecular compounds increased the number of iPSC colonies on both substrates. In the cell proliferation assay, eiPSC-511 showed higher growth activity than eiPSC-111. Although no significant changes were observed in the expression of pluripotency markers between eiPSC-111 and eiPSC-511, the expression of DPPA3 was significantly upregulated in both iPSCs by reprogramming, suggesting that DPPA3 was a sensitive pluripotent marker for equine iPSC. While both iPSCs expressed high mRNA level of integrin alpha6 and beta1 subunits, mRNA level corresponding to ITGA3 and ITGA7 significantly increased in eiPSC-511 in comparison to those in eiPSC-111. These results suggested that the binding strength to the substrate in eiPSC-511 was stronger than that in eiPSC-111. On the contrary, although no significant differences were observed in the histology of teratomas, increased in vitro differentiation into three germ layers in eiPSC-111 was shown compared to those in eiPSC-511. Thus, these results contributed to the improved generation of iPSC in horses.
Copyright © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.
Publication Date: 2025-04-12 PubMed ID: 40228787DOI: 10.1016/j.tvjl.2025.106351Google Scholar: Lookup
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Summary
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The research article focuses on evaluating the effects of different cell adhesion substrates on the generation of equine-induced pluripotent stem cells (eiPSC) from foetal skin cells by using lipofection of plasmid vectors expressing specific reprogramming factors.
Objective of the research
- The main objective of this study was to explore and evaluate the effects of cell adhesion substrates in generating pluripotent stem cells from equine foetal skin cells, specifically focusing on two types of eiPSCs – eiPSC-511 and eiPSC-111.
Findings of the research
- The study found that the reprogramming efficiency of cells on the E8 fragment of laminin-511 (eiPSC-511) was higher than on Geltrex containing laminin-111 as a major laminin (eiPSC-111). This meant that eiPSC-511 cells were more efficient in transforming into pluripotent stem cells than eiPSC-111.
- Supplementation with a mix of small molecular compounds further increased the number of iPSC colonies, indicating that these molecules play a key role in enhancing cell proliferation.
- In cell proliferation assays, eiPSC-511 showed greater growth activity than eiPSC-111. However, there were no significant changes observed in the expression of key pluripotency markers between eiPSC-111 and eiPSC-511, marking their similar pluripotent nature.
- A marked upregulation of DPPA3 expression was observed in both eiPSCs, deducing DPPA3 as a sensitive pluripotent marker for equine iPSC.
- Both iPSCs showed high mRNA levels for integrin alpha6 and beta1 subunits, but the mRNA level corresponding to ITGA3 and ITGA7 increased significantly in eiPSC-511 compared to eiPSC-111. This indicated a stronger binding strength to the substrate in eiPSC-511 than eiPSC-111.
- Despite no significant differences in the histology of teratomas (a type of tumor), eiPSC-111 showed an increased capacity for in vitro differentiation into three germ layers compared to eiPSC-511, which indicates a greater differentiation potential.
Significance of the research
- The boosted generation of eiPSC-511 and its superior binding strength might open new opportunities for their use in advanced therapeutic applications.
- The study provides valuable insights that could contribute to further improving the generation of induced pluripotent stem cells in horses, which might offer potential solutions and treatments to equine diseases.
Cite This Article
APA
Kushida C, Usui T, Tamura N, Kasashima Y, Sato K, Arai K.
(2025).
Comparison of equine-induced pluripotent stem cell characteristics induced on different cell adhesion substrates.
Vet J, 312, 106351.
https://doi.org/10.1016/j.tvjl.2025.106351 Publication
Researcher Affiliations
- Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo, Japan; National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Tokyo, Japan.
- Department of Veterinary Pharmacology, Tokyo University of Agriculture and Technology, Tokyo, Japan.
- Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi, Japan.
- Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi, Japan.
- National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Tokyo, Japan. Electronic address: kota_sato060@maff.go.jp.
- Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo, Japan.
Conflict of Interest Statement
Declaration of Competing Interest The authors declare that they have no competing interests.
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