NAI-CHEN CHENGWang S.Tai-Horng Young2020-03-092020-03-0920120142-9612https://www.scopus.com/inward/record.uri?eid=2-s2.0-83555179053&doi=10.1016%2fj.biomaterials.2011.11.049&partnerID=40&md5=e4f13ad04dd7ad067a14fc737febff9ehttps://scholars.lib.ntu.edu.tw/handle/123456789/474119Adipose-derived stem cells (ASCs) have valuable applications in regenerative medicine, but maintaining the stemness of ASCs during in vitro culture is still a challenging issue. In this study, human ASCs spontaneously formed three-dimensional spheroids on chitosan films. Most ASCs within the spheroid were viable, and the cells produced more extracellular molecules, like laminin and fibronectin. Comparing to monolayer culture, ASC spheroids also exhibited enhanced cell survival in serum deprivation condition. Although cell proliferation was inhibited in spheroids, ASCs readily migrated out and proliferated upon transferring spheroids to another adherent growth surface. Moreover, spheroid-derived ASCs exhibited higher expansion efficiency and colony-forming activity. Importantly, we demonstrated that spheroid formation of human ASCs on chitosan films induced significant upregulation of pluripotency marker genes (Sox-2, Oct-4 and Nanog). By culturing the ASC spheroids in proper induction media, we found that ASC differentiation capabilities were significantly enhanced after spheroid formation, including increased transdifferentiation efficiency into neuron and hepatocyte-like cells. In a nude mice model, we further showed a significantly higher cellular retention ratio of ASC spheroids after intramuscular injection of spheroids and dissociated ASCs. These results suggested that ASCs cultured as spheroids on chitosan films can increase their therapeutic potentials. ? 2011 Elsevier Ltd.[SDGs]SDG3Adipose derived stem cells; Cell survival; Chitosan film; Expansion efficiency; Extracellular; Growth surfaces; In-vitro; Intramuscular injections; Laminin; Marker genes; Monolayer culture; Nude mice; Pluripotency; Regenerative medicine; Retention ratio; Serum deprivation; Spheroid; Stemness; Therapeutic potentials; Transdifferentiation; Up-regulation; Cell proliferation; Chitosan; Differentiation (calculus); Genes; Monolayers; Stem cells; Cell culture; chitosan; octamer transcription factor 4; transcription factor NANOG; transcription factor Sox2; adipose derived stem cell; animal cell; article; cell culture; cell differentiation; cell proliferation; colony forming unit; controlled study; female; in vitro study; liver cell; mouse; nonhuman; priority journal; spheroid cell; upregulation; Adipocytes; Adipose Tissue; Animals; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell Survival; Chitosan; Coculture Techniques; Fibronectins; Gene Expression Regulation; Hepatocytes; Humans; Laminin; Mice; Mice, Nude; Polymerase Chain Reaction; Stem Cells; Mus musculusjournal article10.1016/j.biomaterials.2011.11.049221538702-s2.0-83555179053