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We have used collagenase digestion to isolate the entire limbal epithelial SCs and subjacent mesenchymal cells, and we have demonstrated that their close association is crucial for promoting epithelial clonal growth, implying that the latter serves as niche cells (NCs). After their close association was disrupted by trypsin\/EDTA, single SCs and NCs could reunite to generate sphere growth in three-dimensional Matrigel in the embryonic SC medium, and that such sphere growth initiated by SC\u2013NC reunion was mediated by SDF-1 uniquely expressed by limbal epithelial progenitor cells and its receptor CXCR4, but not CXCR7, strongly expressed by limbal stromal NCs. Inhibition of CXCR4 by AMD3100 or a blocking antibody to CXCR4 but not CXCR7 disrupted their reunion and yielded separate spheres with a reduced size, while resultant epithelial spheres exhibited more corneal differentiation and a notable loss of holoclones. For the first time, these results provide strong evidence supporting that limbal SC function depends on close physical association with their native NCs via SDF-1\/CXCR4 signaling. This novel in vitro model of sphere growth with NCs can be used for investigating how limbal SC self-renewal and fate decision might be regulated in the limbal niche.<\/jats:p>","DOI":"10.1002\/stem.743","type":"journal-article","created":{"date-parts":[[2011,9,24]],"date-time":"2011-09-24T19:39:32Z","timestamp":1316893172000},"page":"1874-1885","source":"Crossref","is-referenced-by-count":93,"title":["Limbal Epithelial Stem\/Progenitor Cells Attract Stromal Niche Cells by SDF-1\/CXCR4 Signaling to Prevent Differentiation"],"prefix":"10.1093","volume":"29","author":[{"given":"Hua-Tao","family":"Xie","sequence":"first","affiliation":[{"name":"R&D Department, Tissue Tech., Inc., Ocular Surface Center, Ocular Surface Research & Education Foundation, Miami, Florida, USA"},{"name":"Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Szu-Yu","family":"Chen","sequence":"additional","affiliation":[{"name":"R&D Department, Tissue Tech., Inc., Ocular Surface Center, Ocular Surface Research & Education Foundation, Miami, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gui-Gang","family":"Li","sequence":"additional","affiliation":[{"name":"R&D Department, Tissue Tech., Inc., Ocular Surface Center, Ocular Surface Research & Education Foundation, Miami, Florida, USA"},{"name":"Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scheffer C.G.","family":"Tseng","sequence":"additional","affiliation":[{"name":"R&D Department, Tissue Tech., Inc., Ocular Surface Center, Ocular Surface Research & Education Foundation, Miami, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2011,10,25]]},"reference":[{"key":"2022011215580615000_bib1","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1038\/35102160","article-title":"Stem cells find their niche","volume":"414","author":"Spradling","year":"2001","journal-title":"Nature"},{"key":"2022011215580615000_bib2","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1016\/S0092-8674(04)00255-7","article-title":"Socializing with the neighbors: Stem cells and their niche","volume":"116","author":"Fuchs","year":"2004","journal-title":"Cell"},{"key":"2022011215580615000_bib3","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1126\/science.1092436","article-title":"Defining the epithelial stem cell niche in skin","volume":"303","author":"Tumbar","year":"2004","journal-title":"Science"},{"key":"2022011215580615000_bib4","doi-asserted-by":"crossref","first-page":"2001","DOI":"10.1242\/dev.002022","article-title":"Stem cells and their niche: An inseparable relationship","volume":"134","author":"Xie","year":"2007","journal-title":"Development"},{"key":"2022011215580615000_bib5","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1083\/jcb.103.1.49","article-title":"Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells","volume":"103","author":"Schermer","year":"1986","journal-title":"J Cell Biol"},{"key":"2022011215580615000_bib6","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1016\/j.exer.2003.09.008","article-title":"Corneal epithelial stem cells at the limbus: Looking at some old problems from a new angle","volume":"78","author":"Lavker","year":"2004","journal-title":"Exp Eye Res"},{"key":"2022011215580615000_bib7","doi-asserted-by":"crossref","first-page":"1476","DOI":"10.1016\/S0161-6420(95)30842-1","article-title":"Cytologic evidence of corneal diseases with limbal stem cell deficiency","volume":"102","author":"Puangsricharern","year":"1995","journal-title":"Ophthalmology"},{"key":"2022011215580615000_bib8","doi-asserted-by":"crossref","first-page":"841","DOI":"10.2174\/156652410793937796","article-title":"Critical appraisal of ex vivo expansion of human limbal epithelial stem cells","volume":"10","author":"Tseng","year":"2010","journal-title":"Curr Mol Med"},{"key":"2022011215580615000_bib9","doi-asserted-by":"crossref","first-page":"4275","DOI":"10.1167\/iovs.03-0089","article-title":"Novel enzymatic isolation of an entire viable human limbal epithelial sheet","volume":"44","author":"Espana","year":"2003","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2022011215580615000_bib10","first-page":"537","article-title":"A new isolation method of human limbal progenitor cells by maintaining close association with their niche cells. 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