Microarray-based gene expression analysis of human osteoblasts in response to different biomaterials

2009 ◽  
Vol 88A (2) ◽  
pp. 401-408 ◽  
Author(s):  
Karina F. Bombonato-Prado ◽  
Larissa S. Bellesini ◽  
Cristina M. Junta ◽  
Márcia M. Marques ◽  
Geraldo A. Passos ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Human Osteoblasts

scholarly journals In Vitro Co-culture Model of Primary Human Osteoblasts and Osteocytes in Collagen Gels

2019 ◽  
Vol 20 (8) ◽  
pp. 1998 ◽  
Author(s):  
Skottke ◽  
Gelinsky ◽  
Bernhardt
Keyword(s):  
Gene Expression ◽  
Bone Tissue ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Porous Membrane ◽  
Collagen Gels ◽  
Rankl Expression ◽  
Human Osteoblasts ◽  
Apical Side

Background: Osteocytes are the key regulator cells in bone tissue, affecting activity of both osteoblasts and osteoclasts. Current in vitro studies on osteocyte-osteoblast interaction are invariably performed with rodent cells, mostly murine cell lines, which diminishes the clinical relevance of the data. Objective: The objective of the present study was to establish an in vitro co-culture system of osteoblasts and osteocytes, which is based solely on human primary cells. Methods: Three different approaches for the generation of human primary osteocytes were compared: direct isolation of osteocytes from bone tissue by multistep digestion, long-time differentiation of human pre-osteoblasts embedded in collagen gels, and short time differentiation of mature human osteoblasts in collagen gels. Co-cultivation of mature osteoblasts with osteocytes, derived from the three different approaches was performed in a transwell system, with osteocytes, embedded in collagen gels at the apical side and osteoblasts on the basal side of a porous membrane, which allowed the separate gene expression analysis for osteocytes and osteoblasts. Fluorescence microscopic imaging and gene expression analysis were performed separately for osteocytes and osteoblasts. Results: All examined approaches provided cells with typical osteocytic morphology, which expressed osteocyte markers E11, osteocalcin, phosphate regulating endopeptidase homolog, X-linked (PHEX), matrix extracellular phosphoglycoprotein (MEPE), sclerostin, and receptor activator of NF-κB Ligand (RANKL). Expression of osteocyte markers was not significantly changed in the presence of osteoblasts. In contrast, osteocalcin gene expression of osteoblasts was significantly upregulated in all examined co-cultures with differentiated osteocytes. Alkaline phosphatase (ALPL), bone sialoprotein II (BSPII), and RANKL expression of osteoblasts was not significantly changed in the co-culture. Conclusion: Interaction of osteoblasts and osteocytes can be monitored in an in vitro model, comprising solely primary human cells.

Sign in / Sign up

Export Citation Format

Share Document