title: Scaffold-free and scaffold-assisted 3D culture enhances differentiation of bone marrow stromal cells.
abstract: 3D cultures of stem cells can preserve differentiation potential or increase the efficiency of methods that induce differentiation. Mouse bone marrow-derived stromal cells (BMSCs) were cultured in 3D as scaffold-free spheroids or "mesoid bodies" (MBs) and as aggregates on poly(lactic) acid microspheres (MB/MS). 3D cultures demonstrated viable cells, interaction on multiple planes, altered cell morphology, and the formation of structures similar to epithelial cell bridges. Cell proliferation was limited in suspension cultures of MB and MB/MS; however, cells regained proliferative capacity when transferred to flat substrates of tissue culture plates (TCPs). Expanded as monolayer, cells retained expression of Sca-1 and CD44 stem cell markers. 3D cultures demonstrated enhanced potential for adipogenic and osteogenic differentiation showing higher triglyceride accumulation and robust mineralization in comparison with TCP cultures. Enhanced and efficient adipogenesis was also observed in 3D cultures generated in a rotating cell culture system. Preservation of multilineage potential of BMSC was demonstrated in 5-azacytidine treatment of 3D cultures and TCP by expression of cardiac markers GATA4 and ACTA1 although functioning cardiomyocytes were not derived.
authors.affiliation: Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600 036, India.
authors.forename: Prasanna
authors.initials: P
authors.lastname: Vidyasekar
authors.affiliation: Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600 036, India.
authors.forename: Pavithra
authors.initials: P
authors.lastname: Shyamsunder
authors.affiliation: Institute of Life Sciences, Nalco Square, Bhubaneswar, 751023, India.
authors.forename: Sanjeeb Kumar
authors.initials: SK
authors.lastname: Sahoo
authors.affiliation: Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600 036, India. vermars@iitm.ac.in.|Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036, Tamil Nadu, India. vermars@iitm.ac.in.
authors.forename: Rama Shanker
authors.initials: RS
authors.lastname: Verma
mesh_terms: D000199:Actins / Q000096:biosynthesis; D050156:Adipogenesis / Q000235:genetics*; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics*; D049109:Cell Proliferation / Q000235:genetics; D050991:GATA4 Transcription Factor / Q000096:biosynthesis; D018507:Gene Expression Regulation, Developmental; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051379:Mice; D032383:Myocytes, Cardiac / Q000378:metabolism; D010012:Osteogenesis / Q000235:genetics*; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: C558396:Acta2 protein, mouse; D000199:Actins; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse
keywords: 3D culture; Adipogenesis; Differentiation; Mesenchymal stem cells; RCCS