Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study

Abstract The scarcity of chondrocytes is a major challenge for cartilage tissue engineering. Monolayer expansion is necessary to amplify the limited number of chondrocytes needed for clinical application. Growth factors are often added to improve monolayer culture conditions, promoting proliferation...

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Autores principales:	Maimonah-Eissa Al-Masawa, Wan Safwani Wan Kamarul Zaman, Kien-Hui Chua
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9b3e3dd6ee074ad39451cc5a24c3fe18
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spelling	oai:doaj.org-article:9b3e3dd6ee074ad39451cc5a24c3fe182021-12-02T12:33:05ZBiosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study10.1038/s41598-020-78395-y2045-2322https://doaj.org/article/9b3e3dd6ee074ad39451cc5a24c3fe182020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78395-yhttps://doaj.org/toc/2045-2322Abstract The scarcity of chondrocytes is a major challenge for cartilage tissue engineering. Monolayer expansion is necessary to amplify the limited number of chondrocytes needed for clinical application. Growth factors are often added to improve monolayer culture conditions, promoting proliferation, and enhancing chondrogenesis. Limited knowledge on the biosafety of the cell products manipulated with growth factors in culture has driven this study to evaluate the impact of growth factor cocktail supplements in chondrocyte culture medium on chondrocyte genetic stability and tumorigenicity. The growth factors were basic fibroblast growth factor (b-FGF), transforming growth factor β2 (TGF β2), insulin-like growth factor 1 (IGF-1), insulin-transferrin-selenium (ITS), and platelet-derived growth factor (PD-GF). Nasal septal chondrocytes cultured in growth factor cocktail exhibited a significantly high proliferative capacity. Comet assay revealed no significant DNA damage. Flow cytometry showed chondrocytes were mostly at G0-G1 phase, exhibiting normal cell cycle profile with no aneuploidy. We observed a decreased tumour suppressor genes’ expression (p53, p21, pRB) and no TP53 mutations or tumour formation after 6 months of implantation in nude mice. Our data suggest growth factor cocktail has a low risk of inducing genotoxic and tumorigenic effects on chondrocytes up to passage 6 with 16.6 population doublings. This preclinical tumorigenicity and genetic instability evaluation is crucial for further clinical works.Maimonah-Eissa Al-MasawaWan Safwani Wan Kamarul ZamanKien-Hui ChuaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maimonah-Eissa Al-Masawa Wan Safwani Wan Kamarul Zaman Kien-Hui Chua Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
description	Abstract The scarcity of chondrocytes is a major challenge for cartilage tissue engineering. Monolayer expansion is necessary to amplify the limited number of chondrocytes needed for clinical application. Growth factors are often added to improve monolayer culture conditions, promoting proliferation, and enhancing chondrogenesis. Limited knowledge on the biosafety of the cell products manipulated with growth factors in culture has driven this study to evaluate the impact of growth factor cocktail supplements in chondrocyte culture medium on chondrocyte genetic stability and tumorigenicity. The growth factors were basic fibroblast growth factor (b-FGF), transforming growth factor β2 (TGF β2), insulin-like growth factor 1 (IGF-1), insulin-transferrin-selenium (ITS), and platelet-derived growth factor (PD-GF). Nasal septal chondrocytes cultured in growth factor cocktail exhibited a significantly high proliferative capacity. Comet assay revealed no significant DNA damage. Flow cytometry showed chondrocytes were mostly at G0-G1 phase, exhibiting normal cell cycle profile with no aneuploidy. We observed a decreased tumour suppressor genes’ expression (p53, p21, pRB) and no TP53 mutations or tumour formation after 6 months of implantation in nude mice. Our data suggest growth factor cocktail has a low risk of inducing genotoxic and tumorigenic effects on chondrocytes up to passage 6 with 16.6 population doublings. This preclinical tumorigenicity and genetic instability evaluation is crucial for further clinical works.
format	article
author	Maimonah-Eissa Al-Masawa Wan Safwani Wan Kamarul Zaman Kien-Hui Chua
author_facet	Maimonah-Eissa Al-Masawa Wan Safwani Wan Kamarul Zaman Kien-Hui Chua
author_sort	Maimonah-Eissa Al-Masawa
title	Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
title_short	Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
title_full	Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
title_fullStr	Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
title_full_unstemmed	Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
title_sort	biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/9b3e3dd6ee074ad39451cc5a24c3fe18
work_keys_str_mv	AT maimonaheissaalmasawa biosafetyevaluationofcultureexpandedhumanchondrocyteswithgrowthfactorcocktailapreclinicalstudy AT wansafwaniwankamarulzaman biosafetyevaluationofcultureexpandedhumanchondrocyteswithgrowthfactorcocktailapreclinicalstudy AT kienhuichua biosafetyevaluationofcultureexpandedhumanchondrocyteswithgrowthfactorcocktailapreclinicalstudy
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Biosafety evaluation of culture-expanded human chondrocytes with growth factor cocktail: a preclinical study

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