{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T09:38:29Z","timestamp":1778751509686,"version":"3.51.4"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,1,28]],"date-time":"2024-01-28T00:00:00Z","timestamp":1706400000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,1,28]],"date-time":"2024-01-28T00:00:00Z","timestamp":1706400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research","award":["18K07544"],"award-info":[{"award-number":["18K07544"]}]},{"name":"Grants-in-Aid for Research on Nervous and Mental Disorders","award":["28-6"],"award-info":[{"award-number":["28-6"]}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["18ek0109239h0002"],"award-info":[{"award-number":["18ek0109239h0002"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Human urine-derived cells (UDCs) are primary cultured cells originating from the upper urinary tract and are known to be multipotent. We previously developed <jats:italic>MYOD1<\/jats:italic>-transduced UDCs (MYOD1-UDCs) as a model recapitulating the pathogenesis of Duchenne muscular dystrophy (DMD) caused by a lack of dystrophin. MYOD1-UDCs also allow evaluation of the efficacy of exon skipping with antisense oligonucleotides. However, despite the introduction of <jats:italic>MYOD1<\/jats:italic>, some MYOD1-UDCs failed to form myotubes, possibly because of heterogeneity among UDCs. Here, we carried out single-cell RNA-sequencing analyses and revealed that CD90\/Thy-1 was highly expressed in a limited subpopulation of UDCs with high myogenic potency. Furthermore, CD90-positive MYOD1-UDCs, but not CD90-negative cells, could form myotubes expressing high levels of myosin heavy chain and dystrophin. Notably, overexpression of CD90 in CD90-negative MYOD1-UDCs did not enhance myogenic differentiation, whereas CD90 suppression in CD90-positive UDCs led to decreased myotube formation and decreased myosin heavy chain expression. CD90 may thus contribute to the fusion of single-nucleated MYOD1-UDCs into myotubes but is not crucial for promoting the expression of late muscle regulatory factors. Finally, we confirmed that CD90-positive MYOD1-UDCs derived from patients with DMD were a valuable tool for obtaining a highly reproducible and stable evaluation of exon skipping using antisense oligonucleotide.<\/jats:p>","DOI":"10.1038\/s41598-024-52530-5","type":"journal-article","created":{"date-parts":[[2024,1,28]],"date-time":"2024-01-28T13:02:06Z","timestamp":1706446926000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Characterization of CD90\/Thy-1 as a crucial molecular signature for myogenic differentiation in human urine-derived cells through single-cell RNA sequencing"],"prefix":"10.1038","volume":"14","author":[{"given":"Katsuhiko","family":"Kunitake","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Norio","family":"Motohashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takafumi","family":"Inoue","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yutaka","family":"Suzuki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshitsugu","family":"Aoki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,1,28]]},"reference":[{"key":"52530_CR1","doi-asserted-by":"publisher","first-page":"4177","DOI":"10.1111\/febs.12267","volume":"280","author":"TA Patridge","year":"2013","unstructured":"Patridge, T. A. The mdx mouse model as a surrogate for Duchenne muscular dystrophy. FEBS J. 280, 4177\u20134186 (2013).","journal-title":"FEBS J."},{"key":"52530_CR2","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.gendis.2014.07.001","volume":"1","author":"D Zhang","year":"2014","unstructured":"Zhang, D., Wei, G., Li, P., Zhou, X. & Zhang, Y. Urine-derived stem cells: A novel and versatile progenitor source for cell-based therapy and regenerative medicine. Genes Dis. 1, 8\u201317 (2014).","journal-title":"Genes Dis."},{"key":"52530_CR3","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1038\/239231a0","volume":"239","author":"GR Sutherland","year":"1972","unstructured":"Sutherland, G. R. & Bain, A. D. Culture of cells from the urine of newborn children. Nature 239, 231 (1972).","journal-title":"Nature"},{"key":"52530_CR4","doi-asserted-by":"publisher","first-page":"1840","DOI":"10.1002\/stem.1424","volume":"31","author":"S Bharadwaj","year":"2013","unstructured":"Bharadwaj, S. et al. Multipotential differentiation of human urine-derived stem cells: Potential for therapeutic applications in urology. Stem Cells 31, 1840\u20131856 (2013).","journal-title":"Stem Cells"},{"key":"52530_CR5","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1002\/bit.21061","volume":"96","author":"S Terstegge","year":"2007","unstructured":"Terstegge, S. et al. Automated maintenance of embryonic stem cell cultures. Biotechnol. Bioeng. 96, 195\u2013201 (2007).","journal-title":"Biotechnol. Bioeng."},{"key":"52530_CR6","doi-asserted-by":"publisher","first-page":"e0125253","DOI":"10.1371\/journal.pone.0125253","volume":"10","author":"J Guan","year":"2015","unstructured":"Guan, J. et al. Human urine derived stem cells in combination with b-TCP can be applied for bone regeneration. PLoS One 10, e0125253 (2015).","journal-title":"PLoS One"},{"key":"52530_CR7","doi-asserted-by":"publisher","first-page":"890574","DOI":"10.3389\/fcell.2022.890574","volume":"10","author":"Y Shi","year":"2022","unstructured":"Shi, Y. et al. Differentiation capacity of human urine-derived stem cells to retain telomerase activity. Front. Cell Dev. Biol 10, 890574 (2022).","journal-title":"Front. Cell Dev. Biol"},{"key":"52530_CR8","doi-asserted-by":"publisher","first-page":"2181","DOI":"10.1016\/j.transproceed.2016.02.078","volume":"48","author":"W He","year":"2016","unstructured":"He, W. et al. Generation of mesenchymal-like stem cells from urine in pediatric patients. Transplant. Proc 48, 2181\u20132185 (2016).","journal-title":"Transplant. Proc"},{"key":"52530_CR9","doi-asserted-by":"publisher","first-page":"2123","DOI":"10.1089\/ten.tea.2010.0637","volume":"17","author":"S Bharadwaj","year":"2011","unstructured":"Bharadwaj, S. et al. Characterization of urine-derived stem cells obtained from upper urinary tract for use in cell-based urological tissue engineering. Tissue Eng. Part A 17, 2123\u20132132 (2011).","journal-title":"Tissue Eng. Part A"},{"key":"52530_CR10","doi-asserted-by":"publisher","first-page":"2080","DOI":"10.1038\/nprot.2012.115","volume":"7","author":"T Zhou","year":"2012","unstructured":"Zhou, T. et al. Generation of human induced pluripotent stem cells from urine samples. Nat. Protoc. 7, 2080\u20132089 (2012).","journal-title":"Nat. Protoc."},{"key":"52530_CR11","doi-asserted-by":"publisher","first-page":"772","DOI":"10.1089\/hum.2016.079","volume":"27","author":"MS Falzarano","year":"2016","unstructured":"Falzarano, M. S. et al. Duchenne muscular dystrophy myogenic cells from urine-derived stem cells recapitulate the dystrophin genotype and phenotype. Hum. Gene Ther. 27, 772\u2013783 (2016).","journal-title":"Hum. Gene Ther."},{"key":"52530_CR12","doi-asserted-by":"publisher","first-page":"3807","DOI":"10.1038\/s41598-019-40421-z","volume":"9","author":"H Takizawa","year":"2019","unstructured":"Takizawa, H. et al. Modelling duchenne muscular dystrophy in MYOD1-converted urine derived cells treated with 3-deazaneplanocin A hydrochloride. Sci. Rep. 9, 3807 (2019).","journal-title":"Sci. Rep."},{"key":"52530_CR13","doi-asserted-by":"publisher","first-page":"712","DOI":"10.1007\/s11427-018-9543-1","volume":"63","author":"AJ Chen","year":"2020","unstructured":"Chen, A. J. et al. Identification and characterization of two morphologically distinct stem cell subpopulations from human urine samples. Sci. China Life Sci. 63, 712\u2013723 (2020).","journal-title":"Sci. China Life Sci."},{"key":"52530_CR14","doi-asserted-by":"publisher","first-page":"928510","DOI":"10.3389\/fcell.2022.928510","volume":"10","author":"P Hu","year":"2022","unstructured":"Hu, P., Leyton, L., Hagood, J. S. & Barker, T. H. Thy-1-integrin interactions in cis and trans mediate distinctive signaling. Front. Cell Dev. Biol. 10, 928510 (2022).","journal-title":"Front. Cell Dev. Biol."},{"key":"52530_CR15","doi-asserted-by":"publisher","first-page":"895","DOI":"10.1681\/ASN.V34895","volume":"3","author":"D Brown","year":"1992","unstructured":"Brown, D. & Waneck, G. L. Glycosyl-phosphatidylinositol-anchored membrane proteins. J. Am. Soc. Nephrol. 3, 895\u2013906 (1992).","journal-title":"J. Am. Soc. Nephrol."},{"key":"52530_CR16","doi-asserted-by":"publisher","first-page":"3581","DOI":"10.4049\/jimmunol.173.6.3581","volume":"173","author":"SMM Haeryfar","year":"2004","unstructured":"Haeryfar, S. M. M. & Hoskin, D. W. Thy-1: More than a mouse pan-T cell marker. J. Immunol. 173, 3581\u20133588 (2004).","journal-title":"J. Immunol."},{"key":"52530_CR17","doi-asserted-by":"publisher","first-page":"921","DOI":"10.1016\/j.bbamcr.2008.10.004","volume":"1793","author":"TH Barker","year":"2009","unstructured":"Barker, T. H. & Hagood, J. S. Getting a grip on Thy-1 signaling. Biochim. Biophys. Acta Mol. Cell Res. 1793, 921\u2013923 (2009).","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"52530_CR18","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/B978-0-12-407695-2.00004-4","volume":"305","author":"R Herrera-Molina","year":"2013","unstructured":"Herrera-Molina, R. et al. Thy-1-interacting molecules and cellular signaling in cis and trans. Int. Rev. Cell Mol. Biol. 305, 163\u2013216 (2013).","journal-title":"Int. Rev. Cell Mol. Biol."},{"key":"52530_CR19","doi-asserted-by":"publisher","first-page":"6689","DOI":"10.1096\/fj.201802224R","volume":"33","author":"A Saalbach","year":"2019","unstructured":"Saalbach, A. & Anderegg, U. Thy-1: More than a marker for mesenchymal stromal cells. FASEB J. 33, 6689\u20136696 (2019).","journal-title":"FASEB J."},{"key":"52530_CR20","doi-asserted-by":"publisher","first-page":"66","DOI":"10.3389\/fcell.2019.00066","volume":"7","author":"C Sauzay","year":"2019","unstructured":"Sauzay, C., Voutetakis, K., Chatziioannou, A., Chevet, E. & Avril, T. CD90\/Thy-1, a cancer-associated cell surface signaling molecule. Front. Cell Dev. Biol. 7, 66 (2019).","journal-title":"Front. Cell Dev. Biol."},{"key":"52530_CR21","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1293\/tox.23.1","volume":"23","author":"T Yuasa","year":"2010","unstructured":"Yuasa, T., Izawa, T., Kuwamura, M. & Yamate, J. Thy-1 expressing mesenchymal cells in rat nephrogenesis in correlation with cells immunoreactive for \u03b1-smooth muscle actin and vimentin. J. Toxicol. Pathol. 23, 1\u201310 (2010).","journal-title":"J. Toxicol. Pathol."},{"key":"52530_CR22","doi-asserted-by":"publisher","first-page":"991","DOI":"10.1016\/j.bbamcr.2006.08.008","volume":"1763","author":"TA Rege","year":"2006","unstructured":"Rege, T. A. & Hagood, J. S. Thy-1, a versatile modulator of signaling affecting cellular adhesion, proliferation, survival, and cytokine\/growth factor responses. Biochim. Biophys. Acta 1763, 991\u2013999 (2006).","journal-title":"Biochim. Biophys. Acta"},{"key":"52530_CR23","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1038\/ki.1996.59","volume":"49","author":"T Sato","year":"1996","unstructured":"Sato, T. et al. Apoptosis of cultured rat glomerular mesangial cells induced by IgG2a monoclonal anti-Thy-1 antibodies. Kidney Int. 49, 403\u2013412 (1996).","journal-title":"Kidney Int."},{"key":"52530_CR24","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1038\/268163a0","volume":"268","author":"JF Lesley","year":"1977","unstructured":"Lesley, J. F. & Lennon, V. A. Transitory expression of Thy-1 antigen in skeletal muscle development. Nature 268, 163\u2013165 (1977).","journal-title":"Nature"},{"key":"52530_CR25","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1038\/289060a0","volume":"289","author":"FS Walsh","year":"1981","unstructured":"Walsh, F. S. & Ritter, M. A. Surface antigen differentiation during human myogenesis in culture. Nature 289, 60\u201364 (1981).","journal-title":"Nature"},{"key":"52530_CR26","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1016\/0006-8993(84)91083-7","volume":"308","author":"CM Booth","year":"1984","unstructured":"Booth, C. M., Brown, M. C., Keynes, R. J. & Barclay, A. N. Muscles of adult rats and mice express the Thy-1 glycoprotein on denervation. Brain Res. 308, 380\u2013382 (1984).","journal-title":"Brain Res."},{"key":"52530_CR27","doi-asserted-by":"publisher","first-page":"767","DOI":"10.1016\/j.devcel.2018.08.006","volume":"46","author":"E Leikina","year":"2018","unstructured":"Leikina, E. et al. Myomaker and myomerger work independently to control distinct steps of membrane remodeling during myoblast fusion. Dev. Cell 46, 767\u2013780 (2018).","journal-title":"Dev. Cell"},{"key":"52530_CR28","doi-asserted-by":"publisher","first-page":"2685","DOI":"10.1242\/dev.01874","volume":"132","author":"SJ Tapscott","year":"2005","unstructured":"Tapscott, S. J. The circuitry of a master switch: Myod and the regulation of skeletal muscle gene transcription. Development 132, 2685\u20132695 (2005).","journal-title":"Development"},{"key":"52530_CR29","doi-asserted-by":"publisher","first-page":"5434","DOI":"10.1073\/pnas.86.14.5434","volume":"86","author":"H Weintraub","year":"1989","unstructured":"Weintraub, H. et al. Activation of muscle-specific genes in pigment, nerve, fat, liver, and fibroblast cell lines by forced expression of MyoD. Proc. Natl. Acad. Sci. USA 86, 5434\u20135438 (1989).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"52530_CR30","doi-asserted-by":"publisher","first-page":"eabc4062","DOI":"10.1126\/sciadv.abc4062","volume":"6","author":"H Zhang","year":"2020","unstructured":"Zhang, H. et al. Human myotube formation is determined by MyoD-myomixer\/myomaker axis. Sci. Adv. 6, eabc4062 (2020).","journal-title":"Sci. Adv."},{"key":"52530_CR31","doi-asserted-by":"publisher","first-page":"1201","DOI":"10.1038\/s41467-020-14766-3","volume":"11","author":"K Van den Berge","year":"2020","unstructured":"Van den Berge, K. et al. Trajectory-based differential expression analysis for single-cell sequencing data. Nat. Commun. 11, 1201 (2020).","journal-title":"Nat. Commun."},{"key":"52530_CR32","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1007\/978-1-0716-2772-3_9","volume":"2587","author":"K Kunitake","year":"2023","unstructured":"Kunitake, K., Sathyaprakash, C., Motohashi, N. & Aoki, Y. Quantitative evaluation of exon skipping in urine-derived cells for duchenne muscular dystrophy. Methods Mol. Biol. 2587, 153\u2013164 (2023).","journal-title":"Methods Mol. Biol."},{"key":"52530_CR33","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1038\/nbt.4096","volume":"36","author":"A Butler","year":"2018","unstructured":"Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411\u2013420 (2018).","journal-title":"Nat. Biotechnol."},{"key":"52530_CR34","doi-asserted-by":"publisher","first-page":"979","DOI":"10.1038\/nmeth.4402","volume":"14","author":"X Qiu","year":"2017","unstructured":"Qiu, X. et al. Reversed graph embedding resolves complex single-cell trajectories. Nat. Methods 14, 979\u2013982 (2017).","journal-title":"Nat. Methods"},{"key":"52530_CR35","doi-asserted-by":"publisher","first-page":"1101","DOI":"10.1038\/nprot.2008.73","volume":"3","author":"TD Schmittgen","year":"2008","unstructured":"Schmittgen, T. D. & Livak, K. J. Analyzing real-time PCR data by the comparative C(T) method. Nat. Protoc. 3, 1101\u20131108 (2008).","journal-title":"Nat. Protoc."},{"key":"52530_CR36","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1093\/jmicro\/dfy015","volume":"67","author":"T Inoue","year":"2018","unstructured":"Inoue, T. TI Workbench, an integrated software package for electrophysiology and imaging. Microscopy (Oxf) 67, 129\u2013143 (2018).","journal-title":"Microscopy (Oxf)"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-52530-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-52530-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-52530-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,31]],"date-time":"2024-01-31T17:08:55Z","timestamp":1706720935000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-024-52530-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,28]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["52530"],"URL":"https:\/\/doi.org\/10.1038\/s41598-024-52530-5","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,28]]},"assertion":[{"value":"16 June 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 January 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 January 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"2329"}}