Methods and insights from the characterization of osteoprogenitor cells of bats (Mammalia: Chiroptera).
Animals; Bone Marrow Cells/cytology; Cell Differentiation/genetics; Cell Proliferation; Cells; Cellular Reprogramming; Chiroptera; Core Binding Factor Alpha 1 Subunit/genetics/metabolism; Cultured; Inbred C57BL; Mice; Osteoblasts/cytology/*metabolism; Osteocalcin/genetics/metabolism; Osteogenesis/*genetics; Real-Time Polymerase Chain Reaction; Stem Cells/*cytology/metabolism; Transcription Factors/genetics/metabolism
Osteoprogenitor cells contribute to the development and maintenance of skeletal tissues. Bats are unique model taxa whose cellular processes are poorly understood, especially in regards to skeletal biology. Forelimb bones of bats, unlike those of terrestrial mammals, bend during flight and function in controlled deformation. As a first step towards understanding the molecular processes governing deposition of this flexible bone matrix, we provide the first method for isolation and differentiation of cell populations derived from the bone marrow and cortical bone of bats, and compare results with those harvested from C57BL/6J mice. Osteogenic capacity of these cells was assessed via absolute quantitative real-time PCR (qPCR) and through quantification of in vitro mineral deposition. Results indicate the differentiated bone cells of bats display significantly lower gene expression of known osteogenic markers (Runt-related transcription factor (RUNX2), osteocalcin (BGLAP) and osterix (SP7)), and deposit a less-mineralized matrix compared with murine controls. By characterizing the in vitro performance of osteoprogenitor cells throughout differentiation and matrix production, this study lays the ground work for in vitro manipulations of bat stem and osteoprogenitor cells and extends our understanding of the cellular diversity across mammals that occupy different habitats.
Ball H C; Moussa F M; Mbimba T; Orman R; Safadi F F; Cooper L N
Stem cell research
2016
2016-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.scr.2016.05.009" target="_blank" rel="noreferrer noopener">10.1016/j.scr.2016.05.009</a>
Osteoactivin induces transdifferentiation of C2C12 myoblasts into osteoblasts.
Alkaline Phosphatase/biosynthesis; Animals; Cell Line; Cell Transdifferentiation/*genetics; Core Binding Factor Alpha 1 Subunit/biosynthesis; Developmental/genetics; Eye Proteins/*genetics/metabolism; Focal Adhesion Protein-Tyrosine Kinases/biosynthesis; Gene Expression Regulation; Humans; Membrane Glycoproteins/*genetics/metabolism; Mice; Myoblasts/cytology/*metabolism; Osteoblasts/cytology/*metabolism
Osteoactivin (OA) is a novel osteogenic factor important for osteoblast differentiation and function. Previous studies showed that OA stimulates matrix mineralization and transcription of osteoblast specific genes required for differentiation. OA plays a role in wound healing and its expression was shown to increase in post fracture calluses. OA expression was reported in muscle as OA is upregulated in cases of denervation and unloading stress. The regulatory mechanisms of OA in muscle and bone have not yet been determined. In this study, we examined whether OA plays a role in transdifferentiation of C2C12 myoblast into osteoblasts. Infected C2C12 with a retroviral vector overexpressing OA under the CMV promoter were able to transdifferentiate from myoblasts into osteoblasts. Immunofluorescence analysis showed that skeletal muscle marker MF-20 was severely downregulated in cells overexpressing OA and contained significantly less myotubes compared to uninfected control. C2C12 myoblasts overexpressing OA showed an increase in expression of bone specific markers such as alkaline phosphatase and alizarin red staining, and also showed an increase in Runx2 protein expression. We also detected increased levels of phosphorylated focal adhesion kinase (FAK) in C2C12 myoblasts overexpressing OA compared to control. Taken together, our results suggest that OA is able to induce transdifferentiation of myoblasts into osteoblasts through increasing levels of phosphorylated FAK.
Sondag Gregory R; Salihoglu Sibel; Lababidi Suzanne L; Crowder Douglas C; Moussa Fouad M; Abdelmagid Samir M; Safadi Fayez F
Journal of cellular physiology
2014
2014-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jcp.24512" target="_blank" rel="noreferrer noopener">10.1002/jcp.24512</a>