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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 K Department of Osteology and Biomechanics, University Medical Center Hamburg-Eppendorf, 22529 Hamburg, Germany. k.jaehn@uke.de.
  • Mason DJ School of Biosciences, Cardiff University, CF10 3AX Cardiff, UK. masondj@cardiff.ac.uk.
  • Ralphs JR School of Biosciences, Cardiff University, CF10 3AX Cardiff, UK. ralphs@cardiff.ac.uk.
  • Evans BAJ School of Medicine, Cardiff University, CF14 4XN Cardiff, UK. evansba@cardiff.ac.uk.
  • Archer CW College of Medicine, Swansea University, SA2 8PP Swansea, UK. c.archer@swansea.ac.uk.
  • Richards RG School of Biosciences, Cardiff University, CF10 3AX Cardiff, UK. geoff.richards@aofoundation.org.
  • Stoddart MJ AO Research Institute Davos, AO Foundation, 7270 Davos, Switzerland. martin.stoddart@aofoundation.org.
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  • 2018-07-04
Published in:
  • International journal of molecular sciences. - 2018
bone biology
osteoblast
osteocyte
three-dimensional (3D) co-culture
transforming growth factor-beta 3 (TGFβ3)
Animals
Cell Death
Cell Differentiation
Cell Survival
Coculture Techniques
Immunohistochemistry
In Situ Nick-End Labeling
Mice
Osteoblasts
Osteocytes
Transforming Growth Factor beta
English 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.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/278774
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