{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T13:44:17Z","timestamp":1791726257441,"version":"4.3.4"},"reference-count":34,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T00:00:00Z","timestamp":1641772800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Health and Welfare","award":["HI16C2207"],"award-info":[{"award-number":["HI16C2207"]}]},{"name":"Ministry of Education","award":["2017M3A9B3063636"],"award-info":[{"award-number":["2017M3A9B3063636"]}]},{"name":"Ministry of Education","award":["2018R1D1A1A09083264"],"award-info":[{"award-number":["2018R1D1A1A09083264"]}]},{"DOI":"10.13039\/501100002630","name":"Ewha Womans University","doi-asserted-by":"publisher","award":["RP-Grant 2020"],"award-info":[{"award-number":["RP-Grant 2020"]}],"id":[{"id":"10.13039\/501100002630","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mesenchymal stem cells (MSCs) can differentiate into endoderm lineages, especially parathyroid-hormone (PTH)-releasing cells. We have previously reported that tonsil-derived MSC (T-MSC) can differentiate into PTH-releasing cells (T-MSC-PTHCs), which restored the parathyroid functions in parathyroidectomy (PTX) rats. In this study, we demonstrate quality optimization by standardizing the differentiation rate for a better clinical application of T-MSC-PTHCs to overcome donor-dependent variation of T-MSCs. Quantitation results of PTH mRNA copy number in the differentiated cells and the PTH concentration in the conditioned medium confirmed that the differentiation efficiency largely varied depending on the cells from each donor. In addition, the differentiation rate of the cells from all the donors greatly improved when differentiation was started at a high cell density (100% confluence). The large-scale expression profiling of T-MSC-PTHCs by RNA sequencing indicated that those genes involved in exiting the differentiation and the cell cycle were the major pathways for the differentiation of T-MSC-PTHCs. Furthermore, the implantation of the T-MSC-PTHCs, which were differentiated at a high cell density embedded in hyaluronic acid, resulted in a higher serum PTH in the PTX model. This standardized efficiency of differentiation into PTHC was achieved by initiating differentiation at a high cell density. Our findings provide a potential solution to overcome the limitations due to donor-dependent variation by establishing a standardized differentiation protocol for the clinical application of T-MSC therapy in treating hypoparathyroidism.<\/jats:p>","DOI":"10.3390\/ijms23020715","type":"journal-article","created":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T17:42:25Z","timestamp":1641836545000},"page":"715","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Density-Dependent Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Parathyroid-Hormone-Releasing Cells"],"prefix":"10.3390","volume":"23","author":[{"given":"Ji Yeon","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saeyoung","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9422-1079","authenticated-orcid":false,"given":"Se-Young","family":"Oh","sequence":"additional","affiliation":[{"name":"Departments of Molecular Medicine, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu Hwa","family":"Nam","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07804, Korea"},{"name":"Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young Min","family":"Choi","sequence":"additional","affiliation":[{"name":"Departments of Molecular Medicine, College of Medicine, Ewha Womans University, Seoul 07804, Korea"},{"name":"Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeonzi","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07804, Korea"},{"name":"Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0561-4792","authenticated-orcid":false,"given":"Ha Yeong","family":"Kim","sequence":"additional","affiliation":[{"name":"Departments of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo Yeon","family":"Jung","sequence":"additional","affiliation":[{"name":"Departments of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2239-0225","authenticated-orcid":false,"given":"Han Su","family":"Kim","sequence":"additional","affiliation":[{"name":"Departments of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6060-6416","authenticated-orcid":false,"given":"Inho","family":"Jo","sequence":"additional","affiliation":[{"name":"Departments of Molecular Medicine, College of Medicine, Ewha Womans University, Seoul 07804, Korea"},{"name":"Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3174-8965","authenticated-orcid":false,"given":"Sung-Chul","family":"Jung","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, Ewha Womans University, Seoul 07804, Korea"},{"name":"Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 07804, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1158\/1055-9965.EPI-08-0960","article-title":"Rising thyroid cancer incidence in the United States by demographic and tumor characteristics, 1980\u20132005","volume":"18","author":"Enewold","year":"2009","journal-title":"Cancer Epidemiol Biomark. Prev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2317","DOI":"10.1002\/jbmr.483","article-title":"Hypoparathyroidism in the adult: Epidemiology, diagnosis, pathophysiology, target-organ involvement, treatment, and challenges for future research","volume":"26","author":"Bilezikian","year":"2011","journal-title":"J. Bone Miner. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1093\/clinchem\/48.10.1731","article-title":"Kinetic analyses of parathyroid hormone clearance as measured by three rapid immunoassays during parathyroidectomy","volume":"48","author":"Bieglmayer","year":"2002","journal-title":"Clin. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1038\/s41523-021-00343-4","article-title":"Evolution of HER2-low expression from primary to recurrent breast cancer","volume":"12","author":"Miglietta","year":"2021","journal-title":"NPJ Breast Cancer"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Park, S., and Jung, S.C. (2021). New Sources, Differentiation, and Therapeutic Uses of Mesenchymal Stem Cells. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms22105288"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"8616","DOI":"10.1038\/srep08616","article-title":"Tonsil-derived mesenchymal stem cells ameliorate CCI4-induced liver fibrosis in mice via autophagy activation","volume":"5","author":"Park","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1111\/joa.12229","article-title":"Characterization of long-term in vitro culture-related alterations of human tonsil-derived mesenchymal stem cells: Role for CCN1 in replicative senescence-associated increase in osteogenic differentiation","volume":"225","author":"Yu","year":"2014","journal-title":"J. Anat."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"106540","DOI":"10.1155\/2015\/106540","article-title":"Immune suppressive effects of tonsil-derived mesenchymal stem cells on mouse bone-marrow-derived dendritic cells","volume":"2015","author":"Park","year":"2015","journal-title":"Stem Cells Int."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.yexcr.2014.06.007","article-title":"Tonsil-derived mesenchymal stem cells alleviate concanavalin A-induced acute liver injury","volume":"326","author":"Ryu","year":"2014","journal-title":"Exp. Cell Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1002\/jcp.24690","article-title":"CCN1 secreted by tonsil-derived mesenchymal stem cells promotes endothelial cell angiogenesis via integrin \u03b1v\u03b23 and AMPK","volume":"230","author":"Park","year":"2015","journal-title":"J. Cell. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"892","DOI":"10.1089\/scd.2014.0384","article-title":"Increased culture density is linked to decelerated proliferation, prolonged G1 phase, and enhanced propensity for differentiation of self-renewing human pluripotent stem cells","volume":"1","author":"Wu","year":"2015","journal-title":"Stem Cells Dev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1016\/j.cell.2008.02.008","article-title":"Differentiation of embryonic stem cells to clinically relevant populations: Lessons from embryonic development","volume":"22","author":"Murry","year":"2008","journal-title":"Cell"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1002\/jcp.21925","article-title":"Human embryonic stem cells are pre-mitotically committed to self-renewal and acquire a lengthened G1 phase upon lineage programming","volume":"222","author":"Becker","year":"2010","journal-title":"J. Cell Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1089\/scd.2008.0337","article-title":"Differentiation of Human Embryonic Stem Cells to a Parathyroid-Like Phenotype","volume":"18","author":"Bingham","year":"2009","journal-title":"Stem Cells Dev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1938","DOI":"10.3945\/jn.111.141762","article-title":"The calcium-sensing receptor affects fat accumulation via effects on antilipolytic pathways in adipose tissue of rats fed low-calcium diets","volume":"141","author":"He","year":"2011","journal-title":"J. Nutr."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"E915","DOI":"10.1152\/ajpendo.00315.2009","article-title":"The calcium-sensing receptor (CaSR) defends against hypercalcemia independently of its regulation of parathyroid hormone secretion","volume":"297","author":"Kantham","year":"2009","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_17","unstructured":"Longo, D.L., Fauci, A., Kasper, D., Hauser, S., Jameson, J., and Loscalzo, J. (2021, November 23). Harrison\u2019s Principles of Internal Medicine, Available online: https:\/\/catalog.nlm.nih.gov\/permalink\/01NLM_INST\/1o1phhn\/alma9914912503406676."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.biomaterials.2015.06.044","article-title":"Differentiated tonsil-derived mesenchymal stem cells embedded in Matrigel restore parathyroid cell functions in rats with parathyroidectomy","volume":"65","author":"Park","year":"2015","journal-title":"Biomaterials"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.actbio.2016.03.003","article-title":"Scaffold-free parathyroid tissue engineering using tonsil-derived mesenchymal stem cells","volume":"15","author":"Park","year":"2016","journal-title":"Acta Biomater."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1002\/(SICI)1097-4644(19991201)75:3<424::AID-JCB8>3.0.CO;2-8","article-title":"Donor variation in the growth properties and osteogenic potential of human marrow stromal cells","volume":"75","author":"Phinney","year":"1999","journal-title":"J. Cell Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1002\/jor.20402","article-title":"Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering","volume":"25","author":"Siddappa","year":"2007","journal-title":"J. Orthop. Res."},{"key":"ref_22","first-page":"1","article-title":"Donor-dependent variation of human umbilical cord blood mesenchymal stem cells in response to hypoxic preconditioning and amelioration of limb ischemia","volume":"50","author":"Kang","year":"2018","journal-title":"Exp. Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3408","DOI":"10.1074\/jbc.273.6.3408","article-title":"Cell density modulates protein-tyrosine phosphorylation","volume":"273","author":"Batt","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1002\/jbmr.5650090703","article-title":"Paracrine interactions among parathyroid cells: Effect of cell density on cell secretion","volume":"9","author":"Sun","year":"1994","journal-title":"J. Bone Miner. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"20130075","DOI":"10.1098\/rsfs.2013.0075","article-title":"The balance between cell cycle arrest and cell proliferation: Control by the extracellular matrix and by contact inhibition","volume":"4","author":"Goldbeter","year":"2014","journal-title":"Interface Focus"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"F485","DOI":"10.1152\/ajprenal.00608.2009","article-title":"Physiology and pathophysiology of the calcium-sensing receptor in the kidney","volume":"298","author":"Riccardi","year":"2010","journal-title":"Am. J. Physiol. Renal Physiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1078\/0344-0338-00360","article-title":"Coexpression of parathyroid hormone and chromogranin A in secondary hyperparathyroidism:a functional marker for secretory activity of hyperplastic nodules","volume":"199","author":"Tanaka","year":"2003","journal-title":"Pathol. Res. Pract."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1002\/jemt.1070320209","article-title":"The biological significance of storage granules in rat parathyroid cells","volume":"32","author":"Setoguti","year":"1995","journal-title":"Microsc. Res. Tech."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1038\/cr.2008.319","article-title":"Adipogenesis licensing and execution are disparately linked to cell proliferation","volume":"19","author":"Guo","year":"2009","journal-title":"Cell Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2961","DOI":"10.1038\/s41467-018-05388-x","article-title":"Contact inhibition controls cell survival and proliferation via YAP\/TAZ-autophagy axis","volume":"9","author":"Pavel","year":"2018","journal-title":"Nat. Com."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1111\/jocd.12237","article-title":"Physiochemical properties and application of hyaluronic acid: A systematic review","volume":"15","author":"Salwowska","year":"2016","journal-title":"J. Cosmet. Dermatol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1001\/jamafacial.2018.2127","article-title":"Assessment of long-term outcomes of soft-tissue augmentation by injecting fibroblasts suspended in hyaluronic acid filler","volume":"21","author":"Moon","year":"2019","journal-title":"JAMA Facial Plast. Surg."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"18529","DOI":"10.18632\/oncotarget.24908","article-title":"Validation and comparison of two NGS assays for the detection of EGFR T790M resistance mutation in liquid biopsies of NSCLC patients","volume":"6","author":"Vollbrecht","year":"2018","journal-title":"Oncotarget"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Jung, S.Y., Kim, H.Y., Park, H.S., Yin, X.Y., Chung, S.M., and Kim, H.S. (2016). Standardization of A Physiologic Hypoparathyroidism Animal Model. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0163911"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/2\/715\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:01:53Z","timestamp":1760364113000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/2\/715"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,10]]},"references-count":34,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,1]]}},"alternative-id":["ijms23020715"],"URL":"https:\/\/doi.org\/10.3390\/ijms23020715","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,10]]}}}