Cell number and chondrogenesis in human mesenchymal stem cell aggregates is affected by the sulfation level of heparin used as a cell coating

• Type of Material: Article
• Authors: Jennifer Lei, Georgia Institute of Technology; Elda Trevino, Emory University; Johnna Temenoff, Emory University
• Language: English
• Date: 2016-07
• Publisher: Wiley: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2016 Wiley Periodicals, Inc.
• Final Published Version (URL): http://dx.doi.org/10.1002/jbm.a.35713
• Title of Journal or Parent Work: Journal of Biomedical Materials Research Part A
• ISSN: 1549-3296
• Volume: 104
• Issue: 7
• Start Page: 1817
• End Page: 1829
• Grant/Funding Information: Contract grant sponsor: NCRR of the NIH; contract grant number: P40RR017447; This work was supported by an NSF DMR (1207045) grant awarded to JST and the GT BioMAT T32 grant (NIH T32EB006343) to ET.; Contract grant sponsor: NSF DMR; contract grant number: 1207045; Contract grant sponsor: BioMAT T32; contract grant number: NIH T32EB006343
• Keywords: heparin; mesenchymal stem cells; cell coatings; growth factors; chondrocytic differentiation
• Research Categories: Engineering, Mechanical; Engineering, Biomedical; Biology, Cell

Abstract

For particular cell-based therapies, it may be required to culture mesenchymal stem cell (MSC) aggregates with growth factors to promote cell proliferation and/or differentiation. Heparin, a negatively charged glycosaminoglycan (GAG) is known to play an important role in sequestration of positively charged growth factors and, when incorporated within cellular aggregates, could be used to promote local availability of growth factors. We have developed a heparin-based cell coating and we believe that the electrostatic interaction between native heparin and the positively charged growth factors will result in (1) higher cell number in response to fibroblast growth factor-2 (FGF-2) and 2) greater chondrogenic differentiation in response to transforming growth factor-β1 (TGF-β1), compared to a desulfated heparin coating. Results revealed that in the presence of FGF-2, by day 14, heparin-coated MSC aggregates increased in DNA content 8.5 ± 1.6 fold compared to day 1, which was greater than noncoated and desulfated heparin-coated aggregates. In contrast, when cultured in the presence of TGF-β1, by day 21, desulfated heparin-coated aggregates upregulated gene expression of collagen II by 86.5 ± 7.5 fold and collagen X by 37.1 ± 4.7 fold, which was higher than that recorded in the noncoated and heparin-coated aggregates. These observations indicate that this coating technology represents a versatile platform to design MSC culture systems with pairings of GAGs and growth factors that can be tailored to overcome specific challenges in scale-up and culture for MSC-based therapeutics.