{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T06:33:47Z","timestamp":1773815627129,"version":"3.50.1"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T00:00:00Z","timestamp":1773792000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Multilineage-differentiating stress-enduring (Muse) cells, a subpopulation of mesenchymal stem cells (MSCs) marked by stage-specific embryonic antigen 3 (SSEA3), exhibit superior regenerative capacity compared to conventional MSCs, including enhanced tissue homing, pluripotency, and paracrine effects. However, their natural scarcity (1\u20135% in MSC populations) limits therapeutic scalability. In this study, we developed a five-compound small molecule method to obtain compound-enriched Muse-like MSCs and assessed their potential use in treating severe veterinary chronic diseases, such as hepatitis and chronic kidney disease (CKD).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Umbilical cord-derived MSCs from cats and dogs were isolated and cultured, using a combination of five small molecules to obtain enriched Muse-like cells. The cultivation process was verified by immunofluorescence and flow cytometry. Differentiation potential of obtained Muse-like cells was evaluated under lineage-specific conditions. These compound-enriched Muse-like MSCs were administered intravenously (2\u202f\u00d7\u202f10\n                      <jats:sup>6<\/jats:sup>\n                      cells\/kg) to a 6-year-old cat with severe hepatitis (twice a week for 2\u202fweeks) and a 16-year-old dog with CKD (weekly for 4\u202fweeks). Serum biochemistry and clinical observations were monitored pre- and post-treatment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Screening of over 100 small-molecule compounds identified optimized five-compound cocktails\u2014valproic acid (0.5\u202fmM), CHIR99021 (3\u202f\u03bcM), PD0325901 (0.5\u202f\u03bcM), Trolox (10\u202f\u03bcM), and nicotinamide (1\u202fmM) for feline MSCs; parnate (10\u202f\u03bcM), CHIR99021 (3\u202f\u03bcM), PD0325901 (0.5\u202f\u03bcM), Trolox (10\u202f\u03bcM), and Y27632 (10\u202f\u03bcM) for canine MSCs. These small-molecule cocktail effectively boosted SSEA3 positivity, from 0.1\u20131% to approximately 40%, as confirmed by immunofluorescence staining and flow cytometry. These enriched Muse-like cells demonstrated superior stress tolerance and robust spontaneous differentiation into endodermal (KRT7\u202f+\u202fhepatocyte-like), mesodermal (cTnI+ cardiomyocyte-like), and ectodermal (Nestin+ neural progenitor) lineages under targeted induction conditions, which were not observed in untreated MSCs. In therapeutic applications, the enriched Muse-like MSCs normalized feline liver indices by day 21 and improved canine renal markers by day 28, accompanied by notable anti-aging effects.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>A small molecule cocktail method was introduced to enhance the Muse population in MSCs, which provide a safe and effective way to harvest Muse cells. These Muse-like MSCs demonstrate high clinical potential for chronic and age-related degenerative diseases, making it a safe and effective therapeutic option.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1708295","type":"journal-article","created":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T05:38:25Z","timestamp":1773812305000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Muse-like stem cell therapy for curing chronic diseases in geriatric feline and canine"],"prefix":"10.3389","volume":"13","author":[{"given":"Yu","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Clinical Research Centre, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine","place":["Shanghai, China"]},{"name":"Shanghai Wholesomebio Technology Co., Ltd.","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wataru","family":"Otsubo","sequence":"additional","affiliation":[{"name":"Shanghai Ainikang Veterinary Company Limited","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aoli","family":"Li","sequence":"additional","affiliation":[{"name":"Shanghai Wholesomebio Technology Co., Ltd.","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hirofumi","family":"Hagino","sequence":"additional","affiliation":[{"name":"Shanghai Ainikang Veterinary Company Limited","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aijie","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haishi","family":"Fan","sequence":"additional","affiliation":[{"name":"Shanghai Wholesomebio Technology Co., Ltd.","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chenwen","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Clinical Research Centre, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine","place":["Shanghai, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,18]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"8639","DOI":"10.1073\/pnas.0911647107","article-title":"Unique multipotent cells in adult human mesenchymal cell populations","volume":"107","author":"Kuroda","year":"2010","journal-title":"Proc Natl Acad Sci 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fibrosis","volume":"26","author":"Iseki","year":"2017","journal-title":"Cell Transplant"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"961","DOI":"10.3390\/cells10040961","article-title":"Non-tumorigenic pluripotent reparative muse cells provide a new therapeutic approach for neurologic diseases","volume":"10","author":"Yamashita","year":"2021","journal-title":"Cells"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1080\/15384101.2016.1211215","article-title":"The secretome of MUSE cells contains factors that may play a role in regulation of stemness, apoptosis and immunomodulation","volume":"16","author":"Alessio","year":"2017","journal-title":"Cell Cycle"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1111\/pin.12129","article-title":"Muse cells, newly found non-tumorigenic pluripotent stem cells, reside in human mesenchymal tissues","volume":"64","author":"Wakao","year":"2014","journal-title":"Pathol 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applications","volume":"11","author":"Ossanna","year":"2023","journal-title":"Biomedicine"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1016\/j.reth.2024.03.003","article-title":"Biological characteristics of muse cells derived from MenSCs and their application in acute liver injury and intracerebral hemorrhage diseases","volume":"27","author":"Li","year":"2024","journal-title":"Regen Ther"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"6699571","DOI":"10.1155\/sci\/6699571","article-title":"Safety and effectiveness of muse cell transplantation in a large-animal model of hepatic fibrosis","volume":"2025","author":"Nishina","year":"2025","journal-title":"Stem Cells Int"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"1027961","DOI":"10.3389\/fphar.2022.1027961","article-title":"Endogenous reparative pluripotent muse cells with a unique immune privilege system: hint at a new strategy for controlling acute and chronic inflammation","volume":"13","author":"Kuroda","year":"2022","journal-title":"Front Pharmacol"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1007\/978-4-431-56847-6_11","volume-title":"Muse Cells: Endogenous Reparative Pluripotent Stem Cells","author":"Uchida","year":"2018"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"1549870","DOI":"10.3389\/fvets.2025.1549870","article-title":"Evaluation of cognitive and mobility function in geriatric dogs following treatment with stem cell and stem cell extracellular vesicles derived from embryonic stem cells: a pilot study","volume":"12","author":"Kim","year":"2025","journal-title":"Front Vet Sci"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"905","DOI":"10.1038\/s42003-025-08333-z","article-title":"Multi-omics analysis of canine aging markers and evaluation of stem cell intervention","volume":"8","author":"Li","year":"2025","journal-title":"Commun Biol"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2026.1708295\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T05:38:27Z","timestamp":1773812307000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2026.1708295\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,18]]},"references-count":35,"alternative-id":["10.3389\/fvets.2026.1708295"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2026.1708295","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,18]]},"article-number":"1708295"}}