Sykes, JG, Kuiper, J, Richardson, JB, Wright, KT and Kuiper, NJ (2018) Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy. Osteoarthritis and Cartilage, 26 (S1). ISSN 1522-9653

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Abstract

Purpose
High hopes have been pinned on regenerative medicine strategies to meet the challenge of preventing the progression to osteoarthritis, in particular autologous chondrocyte implantation (ACI). The loss of chondrocyte phenotype during in vitro monolayer expansion, a step needed to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, we determined whether a shorter and quicker monolayer expansion approach could improve chondrogenic differentiation.

Methods
We compared the effects of two serum types, foetal bovine serum (FBS) and Stemulate™ (a commercial source of human platelet lysate), on the expansion and re-differentiation potential of human chondrocytes isolated from 5 individuals. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were formed and cultured for 4 weeks in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity.

Results
Stemulate™ significantly enhanced proliferation rates (average population doublings; FBS 22.19 d ±6.22 d vs Stemulate™ 12.93 d ±4.64 d). Sulphated glycosaminoglycan (sGAG), total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, spherical and had a more intense staining for toluidine blue, indicating a greater chondrogenic capacity.

Conclusions
Overall we have shown that although Stemulate™ positively influences chondrocyte proliferation, it has a negative effect on chondrogenic differentiation potential. This suggests that Stemulate™ may not be the ideal supplement for the expansion of chondrocytes and to maintain chondrocyte phenotype/genotype, and hence for cell therapies in the treatment of cartilage defects, including ACI.

Item Type: Article
Subjects: R Medicine > RC Internal medicine > RC925 Diseases of the musculoskeletal system
Divisions: Faculty of Medicine and Health Sciences > Institute for Science and Technology in Medicine
Related URLs:
Depositing User: Symplectic
Date Deposited: 23 Oct 2018 13:26
Last Modified: 23 Oct 2018 13:30
URI: http://eprints.keele.ac.uk/id/eprint/5452

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