{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T23:54:19Z","timestamp":1777938859926,"version":"3.51.4"},"reference-count":59,"publisher":"SAGE Publications","issue":"5-6","license":[{"start":{"date-parts":[[2023,3,1]],"date-time":"2023-03-01T00:00:00Z","timestamp":1677628800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Tissue Engineering Part A"],"published-print":{"date-parts":[[2023,3]]},"abstract":"<jats:sec>\n                    <jats:title>Objectives:<\/jats:title>\n                    <jats:p>Successful tissue regeneration requires a clinically viable source of mesenchymal stem cells (MSCs). We explored activin receptor-like kinase (ALK)-5 inhibitors to rapidly derive an MSC-like phenotype with high cartilage forming capacity from a xeno-free human embryonic cell line.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>\n                      Embryonic stem cell (ESC) lines (H9 and HADC100) were treated with the ALK-5 inhibitor SB431542; HADC100 cells were additionally treated with ALK-5 inhibitors SB525334 or GW788388. Cells were then seeded upon human fibronectin in the presence of fibroblast growth factor 2 (FGF2) in a serum-free medium. Flow cytometry was used to assess MSC markers (positive for CD73, CD90, and CD105; negative for CD34 and CD45). Differentiation status was assessed through quantitative polymerase chain reaction. Cartilage forming capacity was determined in high-density pellet cultures, in fibrin gels containing extracellular matrix (fibrin-ECM), and after implantation in\n                      <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                      human osteoarthritic cartilage. Gene expression, histology, and immunostaining were used to assess cartilage phenotype, tissue regeneration, and integration.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      Exposure to all three ALK-5 inhibitors lead to expression of mesodermal gene markers and differentiation into MSC-like cells (embryonic stem cell-derived mesenchymal stem cells [ES-MSCs]) based on surface marker expression. ES-MSC in pellet cultures or in fibrin-ECM gels expressed high levels of chondrogenic genes:\n                      <jats:italic toggle=\"yes\">COL2A1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">ACAN<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">COMP<\/jats:italic>\n                      ; and low levels of\n                      <jats:italic toggle=\"yes\">COL1A1<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">RUNX2<\/jats:italic>\n                      . Cell pellets or fibrin constructs implanted into\n                      <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                      human osteoarthritic cartilage defects produced GAG-rich (safranin O positive) and collagen type II-positive neocartilage tissues that integrated well with native diseased tissue.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions:<\/jats:title>\n                    <jats:p>We developed a protocol for rapid differentiation of xeno-free ESC into MSC-like cells with high cartilage forming capacity with potential for clinical applications.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Impact statement<\/jats:title>\n                    <jats:p>Osteoarthritis (OA) is a common disease resulting in significant disability and no approved disease modifying treatment (other than total joint replacement). Embryonic stem cell-derived cell therapy has the potential to benefit patients with cartilage lesions leading to OA and may prevent or delay the need for total joint replacement.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1089\/ten.tea.2022.0164","type":"journal-article","created":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T04:01:27Z","timestamp":1669953687000},"page":"127-140","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":5,"title":["ALK-5 Inhibitors for Efficient Derivation of Mesenchymal Stem Cells from Human Embryonic Stem Cells"],"prefix":"10.1177","volume":"29","author":[{"given":"Shawn P.","family":"Grogan","sequence":"first","affiliation":[{"name":"Shiley Center for Orthopaedic Research and Education at Scripps Clinic, La Jolla, California, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicholas E.","family":"Glembotski","sequence":"additional","affiliation":[{"name":"Shiley Center for Orthopaedic Research and Education at Scripps Clinic, La Jolla, California, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Darryl D.","family":"D'Lima","sequence":"additional","affiliation":[{"name":"Shiley Center for Orthopaedic Research and Education at Scripps Clinic, La Jolla, California, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2023,3,13]]},"reference":[{"key":"e_1_3_6_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/stcltm\/szac078"},{"key":"e_1_3_6_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-014-1926-5"},{"key":"e_1_3_6_4_2","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tea.2009.0629"},{"key":"e_1_3_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exphem.2007.10.007"},{"key":"e_1_3_6_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2010.11.011"},{"key":"e_1_3_6_7_2","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tea.2012.0751"},{"key":"e_1_3_6_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.25623"},{"key":"e_1_3_6_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/art.27229"},{"key":"e_1_3_6_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scr.2009.05.002"},{"key":"e_1_3_6_11_2","doi-asserted-by":"publisher","DOI":"10.5966\/sctm.2014-0101"},{"key":"e_1_3_6_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3210"},{"key":"e_1_3_6_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1683"},{"key":"e_1_3_6_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scr.2019.101497"},{"key":"e_1_3_6_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-20598-y"},{"key":"e_1_3_6_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1002077107"},{"key":"e_1_3_6_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2013.10.012"},{"key":"e_1_3_6_18_2","doi-asserted-by":"publisher","DOI":"10.5966\/sctm.2011-0022"},{"key":"e_1_3_6_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.34"},{"key":"e_1_3_6_20_2","doi-asserted-by":"publisher","DOI":"10.1002\/stem.569"},{"key":"e_1_3_6_21_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13578-020-00516-x"},{"key":"e_1_3_6_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellsig.2014.06.002"},{"issue":"3","key":"e_1_3_6_23_2","first-page":"361","article-title":"Temporal gene expression and DNA methylation during embryonic stem cell derivation","volume":"20","author":"Samadian A","year":"2018","unstructured":"Samadian A, , Hesaraki M, , Mollamohammadi S, et al. Temporal gene expression and DNA methylation during embryonic stem cell derivation. Cell J, 2018; 20(3):361\u2013368.","journal-title":"Cell J"},{"key":"e_1_3_6_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2008.07.001"},{"key":"e_1_3_6_25_2","doi-asserted-by":"publisher","DOI":"10.1089\/clo.2009.0024"},{"key":"e_1_3_6_26_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.117838"},{"key":"e_1_3_6_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/74447"},{"key":"e_1_3_6_28_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.282.5391.1145"},{"key":"e_1_3_6_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tiv.2010.03.016"},{"key":"e_1_3_6_30_2","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/deh854"},{"key":"e_1_3_6_31_2","doi-asserted-by":"publisher","DOI":"10.1089\/ten.teb.2014.0547"},{"key":"e_1_3_6_32_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0035325"},{"key":"e_1_3_6_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41578-020-0199-8"},{"key":"e_1_3_6_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep18136"},{"key":"e_1_3_6_35_2","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/deq040"},{"key":"e_1_3_6_36_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13287-016-0347-7"},{"key":"e_1_3_6_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scr.2015.12.034"},{"key":"e_1_3_6_38_2","doi-asserted-by":"publisher","DOI":"10.1634\/stemcells.2006-0130"},{"key":"e_1_3_6_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scr.2019.101670"},{"key":"e_1_3_6_40_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13287-017-0561-y"},{"key":"e_1_3_6_41_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.0020161"},{"key":"e_1_3_6_42_2","doi-asserted-by":"publisher","DOI":"10.5966\/sctm.2016-0020"},{"key":"e_1_3_6_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep00455"},{"key":"e_1_3_6_44_2","doi-asserted-by":"publisher","DOI":"10.1080\/03008207.2019.1617280"},{"key":"e_1_3_6_45_2","doi-asserted-by":"publisher","DOI":"10.1053\/joca.2000.0366"},{"key":"e_1_3_6_46_2","doi-asserted-by":"publisher","DOI":"10.1089\/ten.2006.12.2141"},{"key":"e_1_3_6_47_2","doi-asserted-by":"publisher","DOI":"10.1159\/000147272"},{"key":"e_1_3_6_48_2","doi-asserted-by":"publisher","DOI":"10.21769\/BioProtoc.3465"},{"key":"e_1_3_6_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.compbiomed.2011.12.012"},{"key":"e_1_3_6_50_2","doi-asserted-by":"publisher","DOI":"10.1124\/mol.65.3.744"},{"key":"e_1_3_6_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2013.551"},{"issue":"4","key":"e_1_3_6_52_2","first-page":"315","article-title":"Minimal criteria for defining multipotent mesenchymal stromal cells","volume":"8","author":"Dominici M","year":"2006","unstructured":"Dominici M, , Le Blanc K, , Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 2006; 8(4):315\u2013317.","journal-title":"The International Society for Cellular Therapy position statement. Cytotherapy"},{"key":"e_1_3_6_53_2","doi-asserted-by":"publisher","DOI":"10.22203\/eCM.v024a09"},{"key":"e_1_3_6_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2021.01.002"},{"key":"e_1_3_6_55_2","first-page":"4479","article-title":"Clinical development of galunisertib (LY2157299 monohydrate), a small molecule inhibitor of transforming growth factor-beta signaling pathway","volume":"9","author":"Herbertz S","year":"2015","unstructured":"Herbertz S, , Sawyer JS, , Stauber AJ, et al. Clinical development of galunisertib (LY2157299 monohydrate), a small molecule inhibitor of transforming growth factor-beta signaling pathway. Drug Des Devel Ther, 2015; 9:4479\u20134499.","journal-title":"Drug Des Devel Ther"},{"key":"e_1_3_6_56_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13045-021-01053-x"},{"key":"e_1_3_6_57_2","doi-asserted-by":"publisher","DOI":"10.3892\/ijmm.2021.4888"},{"key":"e_1_3_6_58_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2019.07.012"},{"key":"e_1_3_6_59_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2018.10.003"},{"key":"e_1_3_6_60_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2015.01.012"}],"container-title":["Tissue Engineering Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1089\/ten.tea.2022.0164","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/ten.tea.2022.0164","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/ten.tea.2022.0164","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T04:53:58Z","timestamp":1777697638000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/full\/10.1089\/ten.tea.2022.0164"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3]]},"references-count":59,"journal-issue":{"issue":"5-6","published-print":{"date-parts":[[2023,3]]}},"alternative-id":["10.1089\/ten.tea.2022.0164"],"URL":"https:\/\/doi.org\/10.1089\/ten.tea.2022.0164","relation":{},"ISSN":["1937-3341","1937-335X"],"issn-type":[{"value":"1937-3341","type":"print"},{"value":"1937-335X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3]]}}}