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Phenotype and viability of MLO-Y4 cells Is maintained by TGFβ3 in a serum-dependent manner within a 3D-co-culture with MG-63 cells

Jähn, Katharina, Mason, Deborah, Ralphs, Jim, Evans, Bronwen, Archer, Charles, Richards, R. Geoff and Stoddart, Martin 2018. Phenotype and viability of MLO-Y4 cells Is maintained by TGFβ3 in a serum-dependent manner within a 3D-co-culture with MG-63 cells. International Journal of Molecular Sciences 19 (7) , 1932. 10.3390/ijms19071932

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Abstract

The osteocyte network inside the bone matrix is of functional importance and osteocyte cell death is a characteristic feature of pathological bone diseases. Osteocytes have emerged as key regulators of bone tissue maintenance, yet maintaining their phenotype during in vitro culture remains challenging. A 3D co-culture system for osteocytes with osteoblasts was recently presented, enabling the determination of more physiological effects of growth factors on cells in vitro. MLO-Y4 cells were embedded within a type I collagen gel and cultured in the presence of surface MG-63 cells. Co-culture was performed in the presence or absence of TGFβ3. Gene expression by quantitative PCR, protein expression by fluorescent immunohistochemistry and cell viability tests were performed. The 3D co-culture induced cell differentiation of MG-63 cells seen by increased type I collagen and osteocalcin mRNA expression. TGFβ3 maintained osteocyte differentiation of MLO-Y4 cells during co-culture as determined by stable E11 and osteocalcin mRNA expression till day 4. Interestingly, most of the effects of TGFβ3 on co-cultured cells were serum-dependent. Also, TGFβ3 reduced cell death of 3D co-cultured MLO-Y4 cells in a serum-dependent manner. This study shows that 3D co-culture upregulates differentiation of MG-63 cells to a more mature osteoblast-like phenotype; while the addition of TGFβ3 maintained the characteristic MLO-Y4 osteocyte-like phenotype and viability in a serum-dependent manner.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Biosciences
Publisher: MDPI
ISSN: 1422-0067
Date of First Compliant Deposit: 5 July 2018
Date of Acceptance: 28 June 2018
Last Modified: 29 May 2019 23:23
URI: http://orca.cf.ac.uk/id/eprint/112983

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