{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T19:06:16Z","timestamp":1784919976298,"version":"3.55.0"},"reference-count":34,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2023,1,1]],"date-time":"2023-01-01T00:00:00Z","timestamp":1672531200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31972526"],"award-info":[{"award-number":["31972526"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32272848"],"award-info":[{"award-number":["32272848"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012453","name":"China Agricultural Research System","doi-asserted-by":"publisher","award":["CARS-39-03"],"award-info":[{"award-number":["CARS-39-03"]}],"id":[{"id":"10.13039\/501100012453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012453","name":"China Agricultural Research System","doi-asserted-by":"publisher","award":["2020KW-028"],"award-info":[{"award-number":["2020KW-028"]}],"id":[{"id":"10.13039\/501100012453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012453","name":"China Agricultural Research System","doi-asserted-by":"publisher","award":["2022GD-TSLD-46"],"award-info":[{"award-number":["2022GD-TSLD-46"]}],"id":[{"id":"10.13039\/501100012453","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,1,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Hair follicle stem cells (HFSCs) are an important basis for hair follicle morphogenesis and hair cycle growth. This cell type also represents an excellent model for studying the gene function and molecular regulation of the hair growth cycle, including proliferation, differentiation, and apoptosis. Basically, the functional investigation of hair growth-regulating genes demands a sufficient amount of HFSCs. However, efficient propagation of HFSCs in goats is a challenging process under the current culture conditions. Here, we investigated the effect of four components, including the Rho-associated protein kinase (ROCK) inhibitor Y-27632, leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), and vitamin C, on cell growth and pluripotency in the basal culture medium (DMEM\/F12 supplemented with 2% fetal bovine serum). We found that adding Y-27632, LIF, and bFGF independently increased the proliferation and pluripotency of goat HFSCs (gHFSCs), with Y-27632 having the most significant effect (P\u2005&amp;lt;\u20050.001). Fluorescence-activated cell sorting of the cell cycle revealed that Y-27632 promoted gHFSC proliferation by inducing the cell cycle from S to G2\/M phase (P\u2005&amp;lt;\u20050.05). We further demonstrated that gHFSCs displayed superior proliferative capacity, clone-forming ability, and differentiation potential in the combined presence of Y-27632 (10 \u03bcM) and bFGF (10 ng\/mL). We termed this novel culture condition as gHFEM, which stands for goat Hair Follicle Enhanced Medium. Taken together, these results indicate that gHFEM is an optimal condition for in vitro culture of gHFSCs, which will subsequently facilitate the study of HF growth and biology.<\/jats:p>","DOI":"10.1093\/jas\/skad235","type":"journal-article","created":{"date-parts":[[2023,7,11]],"date-time":"2023-07-11T01:30:50Z","timestamp":1689039050000},"source":"Crossref","is-referenced-by-count":6,"title":["Efficient establishment of an optimized culture condition for cashmere goat primary hair follicle stem cells"],"prefix":"10.1093","volume":"101","author":[{"given":"Yan","family":"Li","sequence":"first","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miaohan","family":"Jin","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yawei","family":"Gao","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijin","family":"Lu","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Cao","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"},{"name":"Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, CAS Key Laboratory of Primate Neurobiology, State Key Laboratory of Neuroscience, Chinese Academy of Sciences , Shanghai 200031 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yao","family":"Liu","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulin","family":"Chen","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"},{"name":"Key Laboratory of Livestock Biology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1620-1344","authenticated-orcid":false,"given":"Xiaolong","family":"Wang","sequence":"additional","affiliation":[{"name":"International Joint Agriculture Research Center for Animal Bio-Breeding, Ministry of Agriculture and Rural Affairs\/Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University , Yangling 712100 , China"},{"name":"Key Laboratory of Livestock Biology, Northwest A&F University , Yangling 712100 , China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,7,10]]},"reference":[{"key":"2024080300164293000_CIT0001","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1002\/cm.20472","article-title":"Rho-kinase\/ROCK: a key regulator of the cytoskeleton and cell polarity","volume":"67","author":"Amano","year":"2010","journal-title":"Cytoskeleton"},{"key":"2024080300164293000_CIT0002","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1089\/scd.2016.0234","article-title":"Epidermal growth factor induces proliferation of hair follicle-derived mesenchymal stem cells through epidermal growth factor receptor-mediated activation of ERK and AKT signaling pathways associated with upregulation of cyclin D1 and downregulation of p16","volume":"26","author":"Bai","year":"2017","journal-title":"Stem Cells Dev"},{"key":"2024080300164293000_CIT0003","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1002\/mrd.21021","article-title":"ROCK inhibition enhances the recovery and growth of cryopreserved human embryonic stem cells and human induced pluripotent stem cells","volume":"76","author":"Claassen","year":"2009","journal-title":"Mol. Reprod. Dev"},{"key":"2024080300164293000_CIT0035","doi-asserted-by":"crossref","first-page":"3003\u2013","DOI":"10.4161\/cc.21088","article-title":"Cell cycle regualtion of Rho signaling pathways","volume":"11","author":"David","year":"2012","journal-title":"Cell Cycle"},{"key":"2024080300164293000_CIT0004","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.stem.2009.12.001","article-title":"Vitamin C enhances the generation of mouse and human induced pluripotent stem cells","volume":"6","author":"Esteban","year":"2010","journal-title":"Cell Stem Cell"},{"key":"2024080300164293000_CIT0005","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1007\/s00441-022-03583-3","article-title":"Culture supplementation of bFGF, GDNF, and LIF alters in vitro proliferation, colony formation, and pluripotency of neonatal porcine germ cells","volume":"388","author":"Fayaz","year":"2022","journal-title":"Cell Tissue Res"},{"key":"2024080300164293000_CIT0006","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1194\/jlr.M032284","article-title":"Rho\/ROCK pathway is essential to the expansion, differentiation, and morphological rearrangements of human neural stem\/progenitor cells induced by lysophosphatidic acid","volume":"54","author":"Frisca","year":"2013","journal-title":"J. Lipid Res"},{"key":"2024080300164293000_CIT0007","doi-asserted-by":"crossref","first-page":"2579","DOI":"10.1007\/s10616-016-9981-2","article-title":"Isolation and characterization of hair follicle stem cells from Arbas Cashmere goat","volume":"68","author":"He","year":"2016","journal-title":"Cytotechnology"},{"key":"2024080300164293000_CIT0008","doi-asserted-by":"crossref","first-page":"2518","DOI":"10.1128\/MCB.01308-09","article-title":"Runx1 directly promotes proliferation of hair follicle stem cells and epithelial tumor formation in mouse skin","volume":"30","author":"Hoi","year":"2010","journal-title":"Mol. Cell. Biol"},{"key":"2024080300164293000_CIT0009","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1038\/ng.239","article-title":"Lgr5 marks cycling, yet long-lived, hair follicle stem cells","volume":"40","author":"Jaks","year":"2008","journal-title":"Nat. Genet"},{"key":"2024080300164293000_CIT0010","first-page":"356","article-title":"Effect of fasudil hydrochloride on the post-thaw viability of cryopreserved porcine adipose-derived stem cells","volume":"35","author":"Ji","year":"2014","journal-title":"Cryo Letters"},{"key":"2024080300164293000_CIT0011","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1038\/s41577-018-0066-7","article-title":"Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer","volume":"18","author":"Jones","year":"2018","journal-title":"Nat. Rev. Immunol"},{"key":"2024080300164293000_CIT0012","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.neulet.2014.07.011","article-title":"Promotion of mouse embryonic stem cell differentiation by Rho kinase inhibitor Y-27632","volume":"579","author":"Kamishibahara","year":"2014","journal-title":"Neurosci. Lett"},{"key":"2024080300164293000_CIT0013","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1242\/dev.044123","article-title":"Hair follicle renewal: authentic morphogenesis that depends on a complex progression of stem cell lineages","volume":"137","author":"Legu\u00e9","year":"2010","journal-title":"Development"},{"key":"2024080300164293000_CIT0014","doi-asserted-by":"crossref","first-page":"e104365","DOI":"10.15252\/embj.2019104365","article-title":"SIRT7 activates quiescent hair follicle stem cells to ensure hair growth in mice","volume":"39","author":"Li","year":"2020","journal-title":"EMBO J"},{"key":"2024080300164293000_CIT0015","doi-asserted-by":"crossref","first-page":"26120","DOI":"10.1074\/jbc.M212082200","article-title":"Epidermal growth factor as a biologic switch in hair growth cycle","volume":"278","author":"Mak","year":"2003","journal-title":"J. Biol. 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Sci"},{"key":"2024080300164293000_CIT0019","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1038\/sj.cr.7290134","article-title":"Stem cell pluripotency and transcription factor Oct4","volume":"12","author":"Pan","year":"2002","journal-title":"Cell Res"},{"key":"2024080300164293000_CIT0020","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1038\/sj.cr.7310125","article-title":"Nanog and transcriptional networks in embryonic stem cell pluripotency","volume":"17","author":"Pan","year":"2007","journal-title":"Cell Res"},{"key":"2024080300164293000_CIT0021","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.semcdb.2013.12.005","article-title":"Stem cell dynamics in the hair follicle niche","volume":"25-26","author":"Rompolas","year":"2014","journal-title":"Semin. Cell Dev. 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Biol"},{"key":"2024080300164293000_CIT0029","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/nature14413","article-title":"An alternative pluripotent state confers interspecies chimaeric competency","volume":"521","author":"Wu","year":"2015","journal-title":"Nature"},{"key":"2024080300164293000_CIT0030","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.theriogenology.2015.09.020","article-title":"ROCK inhibitor Y27632 promotes proliferation and diminishes apoptosis of marmoset induced pluripotent stem cells by suppressing expression and activity of caspase3","volume":"85","author":"Wu","year":"2016","journal-title":"Theriogenology"},{"key":"2024080300164293000_CIT0031","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1002\/stem.3345","article-title":"LIF maintains mouse embryonic stem cells pluripotency by modulating TET1 and JMJD2 activity in a JAK2-dependent manner","volume":"39","author":"Wulansari","year":"2021","journal-title":"Stem Cells"},{"key":"2024080300164293000_CIT0032","doi-asserted-by":"crossref","first-page":"1368","DOI":"10.7150\/ijbs.33233","article-title":"Exosomal Micro RNAs derived from dermal papilla cells mediate hair follicle stem cell proliferation and differentiation","volume":"15","author":"Yan","year":"2019","journal-title":"Int. J. Biol. Sci"},{"key":"2024080300164293000_CIT0033","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1186\/s12917-022-03420-3","article-title":"Neural, adipocyte and hepatic differentiation potential of primary and secondary hair follicle stem cells isolated from Arbas Cashmere goats","volume":"18","author":"Yan","year":"2022","journal-title":"BMC Vet. 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