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Open AccessCommunication

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

1
Department of Osteology and Biomechanics, University Medical Center Hamburg-Eppendorf, 22529 Hamburg, Germany
2
School of Biosciences, Cardiff University, CF10 3AX Cardiff, UK
3
School of Medicine, Cardiff University, CF14 4XN Cardiff, UK
4
College of Medicine, Swansea University, SA2 8PP Swansea, UK
5
AO Research Institute Davos, AO Foundation, 7270 Davos, Switzerland
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(7), 1932; https://doi.org/10.3390/ijms19071932
Received: 9 May 2018 / Revised: 15 June 2018 / Accepted: 28 June 2018 / Published: 30 June 2018
(This article belongs to the Special Issue Cell-Biomaterial Interaction)
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. View Full-Text
Keywords: bone biology; osteoblast; osteocyte; three-dimensional (3D) co-culture; transforming growth factor-beta 3 (TGFβ3) bone biology; osteoblast; osteocyte; three-dimensional (3D) co-culture; transforming growth factor-beta 3 (TGFβ3)
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MDPI and ACS Style

Jähn, K.; Mason, D.J.; Ralphs, J.R.; Evans, B.A.; Archer, C.W.; Richards, R.G.; Stoddart, M.J. 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. Int. J. Mol. Sci. 2018, 19, 1932.

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