Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 19, Issue 7, Pages -Publisher
MDPI
DOI: 10.3390/ijms19071932
Keywords
bone biology; osteoblast; osteocyte; three-dimensional (3D) co-culture; transforming growth factor-beta 3 (TGF beta(3))
Funding
- AO Foundation
- PIER [PIF-2017-69]
- DFG [JA 2654/1-1]
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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 TGF beta(3). 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. TGF beta(3) 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 TGF beta(3) on co-cultured cells were serum-dependent. Also, TGF beta(3) 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 TGF beta(3) maintained the characteristic MLO-Y4 osteocyte-like phenotype and viability in a serum-dependent manner.
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