title: Spheroid model for functional osteogenic evaluation of human adipose derived stem cells.
abstract: 3D culture systems have the ability to mimic the natural microenvironment by allowing better cell-cell interactions. We have prepared an in vitro 3D osteogenic cell culture model using human adipose derived stem cells (hASCs) cultured atop recombinant elastin-like polypeptide (ELP) conjugated to a charged polyelectrolyte, polyethyleneimine (PEI). We demonstrate that hASCs cultured atop the ELP-PEI coated tissue culture polystyrene (TCPS) formed 3D spheroids and exhibited superior differentiation toward osteogenic lineage compared to the traditional two dimensional (2D) monolayer formed atop uncoated TCPS. Live/dead viability assay confirmed >90% live cells at the end of the 3-week culture period. Over the same culture period, higher protein content was observed in 2D monolayer than 3D spheroids, as the 2D environment allowed continued proliferation, while 3D spheroids underwent contact-inhibited growth arrest. The normalized alkaline phosphatase (ALP) activity, which is an indicator for early osteogenic differentiation was higher for 3D spheroids. The normalized osteocalcin (OCN) production, which is an indicator for osteogenic maturation was also higher for 3D spheroids while 2D monolayer had no noticeable OCN production. On day 22, increased Alizarin red uptake by 3D spheroids showed greater mineralization activity than 2D monolayer. Taken together, these results indicate a superior osteogenic differentiation of hASCs in 3D spheroid culture atop ELP-PEI coated TCPS surfaces than the 2D monolayer formed on uncoated TCPS surfaces. Such enhanced osteogenesis in 3D spheroid stem cell culture may serve as an alternative to 2D culture by providing a better microenvironment for the enhanced cellular functions and interactions in bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1230-1236, 2017.
authors.affiliation: Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, Jackson, Mississippi, 39216.
authors.forename: Bhuvaneswari
authors.initials: B
authors.lastname: Gurumurthy
authors.affiliation: Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, Jackson, Mississippi, 39216.
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Bierdeman
authors.affiliation: Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, Jackson, Mississippi, 39216.
authors.forename: Amol V
authors.initials: AV
authors.lastname: Janorkar
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000328:Adult; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004549:Elastin / Q000737:chemistry; D005260:Female; D006801:Humans; D008954:Models, Biological; D015675:Osteocalcin / Q000096:biosynthesis; D010012:Osteogenesis; D011137:Polystyrenes / Q000737:chemistry; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism*; D055153:Stem Cell Niche; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: D020099:Coated Materials, Biocompatible; D011137:Polystyrenes; D015675:Osteocalcin; D004549:Elastin
keywords: 3D spheroids; bone tissue engineering; elastin-like polypeptide; in vitro 3D model; osteogenesis