title: Temporal profiling of the growth and multi-lineage potentiality of adipose tissue-derived mesenchymal stem cells cell-sheets.
abstract: Cell-sheet tissue engineering retains the benefits of an intact extracellular matrix (ECM) and can be used to produce scaffold-free constructs. Adipose tissue-derived stem cells (ASCs) are multipotent and more easily obtainable than the commonly used bone marrow-derived stem cells (BMSCs). Although BMSC cell sheets have been previously reported to display multipotentiality, a detailed study of the development and multilineage potential of ASC cell sheets (ASC-CSs) is non-existent in the literature. The aims of this study were to temporally profile: (a) the effect of hyperconfluent culture duration on ASC-CSs development; and (b) the multipotentiality of ASC-CSs by differentiation into the osteogenic, adipogenic and chondrogenic lineages. Rabbit ASCs were first isolated and cultured until confluence (day 0). The confluent cells were then cultured in ascorbic acid-supplemented medium for 3 weeks to study cell metabolic activity, cell sheet thickness and early differentiation gene expressions at weekly time points. ASC-CSs and ASCs were then differentiated into the three lineages, using established protocols, and assessed by RT-PCR and histology at multiple time points. ASC-CSs remained healthy up to 3 weeks of hyperconfluent culture. One week-old cell sheets displayed upregulation of early differentiation gene markers (Runx2 and Sox9); however, subsequent differentiation results indicated that they did not necessarily translate to an improved phenotype. ASCs within the preformed cell sheet groups did not differentiate as efficiently as the non-hyperconfluent ASCs, which were directly differentiated. Although ASCs within the cell sheets retained their differentiation capacity and remained viable under prolonged hyperconfluent conditions, future applications of ASC-CSs in tissue engineering should be considered with care. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Bioengineering, Faculty of Engineering, National University of Singapore.
authors.forename: Puay Yong
authors.initials: PY
authors.lastname: Neo
authors.affiliation: Department of Bioengineering, Faculty of Engineering, National University of Singapore.
authors.forename: Eugene Yong-Shun
authors.initials: EY
authors.lastname: See
authors.affiliation: Department of Bioengineering, Faculty of Engineering, National University of Singapore.|Department of Mechanical Engineering, Faculty of Engineering, National University of Singapore.
authors.forename: Siew Lok
authors.initials: SL
authors.lastname: Toh
authors.affiliation: Department of Bioengineering, Faculty of Engineering, National University of Singapore.|Department of Orthopaedic Surgery, Yong Lin Loo School of Medicine, National University of Singapore.
authors.forename: James Cho-Hong
authors.initials: JC
authors.lastname: Goh
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis; D002454:Cell Differentiation; D050659:Core Binding Factor Alpha 1 Subunit / Q000096:biosynthesis; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D011817:Rabbits; D055758:SOX9 Transcription Factor / Q000096:biosynthesis; D015854:Up-Regulation
chemical_list: D000943:Antigens, Differentiation; D050659:Core Binding Factor Alpha 1 Subunit; D055758:SOX9 Transcription Factor
keywords: adipose-tissue derived stem cells; cell sheet; hyperconfluent culture; multi-lineage differentiation; temporal profiling; tissue engineering