Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins

Introduction: Mesenchymal stem cells (MSCs) possess immunomodulatory potential and are used for cell therapy in both human and veterinary medicine. However, donor-derived MSCs have limited proliferative activities and variations, which restrict their clinical applicability. In contrast, induced plur...

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Main Authors: Masaya Tsukamoto, Chiaki Kawabata, Kohei Shishida, Takumi Yoshida, Kazuto Kimura, Kazuya Edamura, Kikuya Sugiura, Shingo Hatoya
Format: Article
Language:English
Published: Elsevier 2025-12-01
Series:Regenerative Therapy
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352320425001245
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author Masaya Tsukamoto
Chiaki Kawabata
Kohei Shishida
Takumi Yoshida
Kazuto Kimura
Kazuya Edamura
Kikuya Sugiura
Shingo Hatoya
author_facet Masaya Tsukamoto
Chiaki Kawabata
Kohei Shishida
Takumi Yoshida
Kazuto Kimura
Kazuya Edamura
Kikuya Sugiura
Shingo Hatoya
author_sort Masaya Tsukamoto
collection DOAJ
description Introduction: Mesenchymal stem cells (MSCs) possess immunomodulatory potential and are used for cell therapy in both human and veterinary medicine. However, donor-derived MSCs have limited proliferative activities and variations, which restrict their clinical applicability. In contrast, induced pluripotent stem cells (iPSCs) can self-renew indefinitely and differentiate into the three germ layers. By exploiting these characteristics, iPSCs can differentiate into mesenchymal stem cells (iMSCs) and potentially overcome the limitations of donor-derived MSCs. In humans, the characteristics of iMSCs have been reported to vary depending on the differentiation strategy and cell origin of iPSCs. However, no studies have investigated the differentiation strategies and cell origins of canine iPSCs (ciPSCs) in relation to iMSC generation. Methods: Canine embryonic fibroblast-derived iPSCs (CEF-iPSCs) were differentiated into iMSCs via the mesoderm or ectoderm, and their proliferative ability and the expression levels of CD34, CD44, CD45, and CD90 were assessed. We then applied the iMSC induction method via the ectoderm to other ciPSC lines, including canine dermal fibroblast-derived iPSCs (CDF-iPSCs), canine peripheral mononuclear cell-derived iPSCs (cPBMC-iPSCs), and canine urine-derived cell-derived iPSCs (cUC-iPSCs). We assessed their proliferation, marker expression, and ability to differentiate into tri-lineages and performed comparative analyses. Results: IMSCs derived from CEF-iPSCs via the ectodermal lineage showed higher proliferative ability and expressed MSC markers at a higher rate than iMSCs generated via mesodermal induction. Notably, with the exception of CDF-iPSCs, iMSCs were successfully generated from other ciPSC lines via ectodermal lineages. These iMSCs exhibited proliferative activities over passage 10, expressed MSC markers, and demonstrated the ability to differentiate into tri-lineages. iMSCs derived from cUC-iPSCs exhibited the highest expression of CD90 compared to other iMSCs. Conclusions: Highly proliferative iMSCs expressing a high rate of MSC markers can be obtained from cUC-iPSCs via ectodermal induction. Our study demonstrated that the differentiation strategy and cell origin of ciPSCs play crucial roles in the generation of iMSCs.
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spelling doaj-art-3ec0b2806c62443eaf47b9cbae4b66492025-08-20T02:34:53ZengElsevierRegenerative Therapy2352-32042025-12-013011212210.1016/j.reth.2025.05.008Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell originsMasaya Tsukamoto0Chiaki Kawabata1Kohei Shishida2Takumi Yoshida3Kazuto Kimura4Kazuya Edamura5Kikuya Sugiura6Shingo Hatoya7Department of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, Japan; Department of Pathology, Microbiology & Immunology, School of Veterinary Medicine, University of California, Davis, CA, USALaboratory of Veterinary Surgery, Department of Veterinary Medicine, College of Bioresource and Sciences, Nihon University, Fujisawa, Kanagawa 252-0880, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, JapanDepartment of Advanced Pathobiology, Graduate School of Veterinary Science, Osaka Metropolitan University, Izumisano, Osaka 598-8531, Japan; Corresponding author.Introduction: Mesenchymal stem cells (MSCs) possess immunomodulatory potential and are used for cell therapy in both human and veterinary medicine. However, donor-derived MSCs have limited proliferative activities and variations, which restrict their clinical applicability. In contrast, induced pluripotent stem cells (iPSCs) can self-renew indefinitely and differentiate into the three germ layers. By exploiting these characteristics, iPSCs can differentiate into mesenchymal stem cells (iMSCs) and potentially overcome the limitations of donor-derived MSCs. In humans, the characteristics of iMSCs have been reported to vary depending on the differentiation strategy and cell origin of iPSCs. However, no studies have investigated the differentiation strategies and cell origins of canine iPSCs (ciPSCs) in relation to iMSC generation. Methods: Canine embryonic fibroblast-derived iPSCs (CEF-iPSCs) were differentiated into iMSCs via the mesoderm or ectoderm, and their proliferative ability and the expression levels of CD34, CD44, CD45, and CD90 were assessed. We then applied the iMSC induction method via the ectoderm to other ciPSC lines, including canine dermal fibroblast-derived iPSCs (CDF-iPSCs), canine peripheral mononuclear cell-derived iPSCs (cPBMC-iPSCs), and canine urine-derived cell-derived iPSCs (cUC-iPSCs). We assessed their proliferation, marker expression, and ability to differentiate into tri-lineages and performed comparative analyses. Results: IMSCs derived from CEF-iPSCs via the ectodermal lineage showed higher proliferative ability and expressed MSC markers at a higher rate than iMSCs generated via mesodermal induction. Notably, with the exception of CDF-iPSCs, iMSCs were successfully generated from other ciPSC lines via ectodermal lineages. These iMSCs exhibited proliferative activities over passage 10, expressed MSC markers, and demonstrated the ability to differentiate into tri-lineages. iMSCs derived from cUC-iPSCs exhibited the highest expression of CD90 compared to other iMSCs. Conclusions: Highly proliferative iMSCs expressing a high rate of MSC markers can be obtained from cUC-iPSCs via ectodermal induction. Our study demonstrated that the differentiation strategy and cell origin of ciPSCs play crucial roles in the generation of iMSCs.http://www.sciencedirect.com/science/article/pii/S2352320425001245CanineInduced pluripotent stem cellMesenchymal stem cellNeural crest cellVeterinary regenerative medicine
spellingShingle Masaya Tsukamoto
Chiaki Kawabata
Kohei Shishida
Takumi Yoshida
Kazuto Kimura
Kazuya Edamura
Kikuya Sugiura
Shingo Hatoya
Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
Regenerative Therapy
Canine
Induced pluripotent stem cell
Mesenchymal stem cell
Neural crest cell
Veterinary regenerative medicine
title Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
title_full Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
title_fullStr Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
title_full_unstemmed Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
title_short Generation of canine induced pluripotent stem cell-derived mesenchymal stem cells: Comparison of differentiation strategies and cell origins
title_sort generation of canine induced pluripotent stem cell derived mesenchymal stem cells comparison of differentiation strategies and cell origins
topic Canine
Induced pluripotent stem cell
Mesenchymal stem cell
Neural crest cell
Veterinary regenerative medicine
url http://www.sciencedirect.com/science/article/pii/S2352320425001245
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