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Home / Equine Research Bank / PLoS One / Calcium-sensing receptor-mediated osteogenic and early-stage...
PloS one2014; 9(11); e111533; doi: 10.1371/journal.pone.0111533

Calcium-sensing receptor-mediated osteogenic and early-stage neurogenic differentiation in umbilical cord matrix mesenchymal stem cells from a large animal model.

Abstract: Umbilical cord matrix mesenchymal stem cells (UCM-MSCs) present a wide range of potential therapeutical applications. The extracellular calcium-sensing receptor (CaSR) regulates physiological and pathological processes. We investigated, in a large animal model, the involvement of CaSR in triggering osteogenic and neurogenic differentiation of two size-sieved UCM-MSC lines, by using AMG641, a novel potent research calcimimetic acting as CaSR agonist. Results: Large (>8 µm in diameter) and small (<8 µm) equine UCM-MSC lines were cultured in medium with high calcium (Ca2+) concentration ([Ca2+]o; 2.87 mM) and dose-response effects of AMG641 (0.01 to 3µM) on cell proliferation were evaluated. Both cell lines were then cultured in osteogenic or neurogenic differentiation medium containing: 1) low [Ca2+]o (0.37 mM); 2) high [Ca2+]o (2.87 mM); 3) AMG641 (0.05, 0.1 or 1 µM) with high [Ca2+]o and 4) the CaSR antagonist NPS2390 (10 mM for 30 min) followed by incubation with AMG641 in high [Ca2+]o. Expression of osteogenic or neurogenic differentiation biomarkers was compared among groups. In both cell lines, AMG641 dose-dependently increased cell proliferation (up to P<0.001). Osteogenic molecular markers expression was differentially regulated by AMG641, with stimulatory (OPN up-regulation) in large or inhibitory (RUNX2 and OPN down-regulation) effects in small cells, respectively. AMG641 significantly increased alkaline phosphatase activity and calcium phosphate deposition in both cell lines. Following treatment with AMG641 during osteogenic differentiation, in both cell lines CaSR expression was inversely related to that of osteogenic markers and inhibition of CaSR by NPS2390 blocked AMG641-dependent responses. Early-stage neurogenic differentiation was promoted/triggered by AMG641 in both cell lines, as Nestin and CaSR mRNA transcription up-regulation were observed. Conclusions: Calcium- and AMG641-induced CaSR stimulation promoted in vitro proliferation and osteogenic and early-stage neurogenic differentiation of UCM-MSCs. CaSR activation may play a fundamental role in selecting specific differentiation checkpoints of these two differentiation routes, as related to cell commitment status.
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Publication Date: 2014-11-07 PubMed ID: 25379789PubMed Central: PMC4224416DOI: 10.1371/journal.pone.0111533Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates how the calcium-sensing receptor plays a role in initiating bone and early nerve cell development in umbilical cord matrix mesenchymal stem cells, using a new research compound that acts as an agonist for this receptor.

Background and Aim of the Study

  • The study aimed to understand the role of the Calcium-sensing receptor (CaSR) in initiating osteogenic (bone cell forming) and early stages of neurogenic (nerve cell forming) differentiation in two size-sieved umbilical cord matrix mesenchymal stem cells (UCM-MSCs) lines in a large animal model.
  • The research team used a new potent research compound, AMG641, that acts as an agonist for the CaSR.

Methods and Procedures

  • Two lines of equine UCM-MSCs were used which ranged in size – large cells greater than 8 µm in diameter, and small cells smaller than 8 µm.
  • These cell lines were cultured in medium with high calcium (Ca2+) concentration and the effect of AMG641 (at various concentrations) on cell proliferation was examined.
  • Calcium concentrations, AMG641 doses, and the use of a CaSR antagonist were varied during culture to observe the responses on cell proliferation and osteogenic or neurogenic differentiation.

Results and Findings

  • The research found that AMG641 dose-dependently increased cell proliferation.
  • AMG641 selectively regulated the expression of osteogenic molecular markers, having a stimulating effect in large cells and an inhibitory effect in small cells.
  • AMG641 also significantly increased alkaline phosphatase activity and calcium phosphate deposition in both cell lines suggesting an enhanced differentiation into bone cells.
  • The expression of the CaSR was inversely related to the expression of osteogenic markers, indicating a regulatory role for CaSR in osteogenic differentiation. When this receptor was blocked, the AMG641-dependent responses were also blocked.
  • Early-stage neurogenic differentiation was promoted by AMG641 in both large and small cell lines.

Conclusion

  • The study concluded that the stimulation of the CaSR by calcium and AMG641 promoted in vitro proliferation and osteogenic and early-stage neurogenic differentiation of UCM-MSCs.
  • CaSR activation may be crucial in choosing specific differentiation checkpoints along the osteogenic and neurogenic differentiation pathways, depending on the cell’s commitment status.

Cite This Article

APA
(2014). Calcium-sensing receptor-mediated osteogenic and early-stage neurogenic differentiation in umbilical cord matrix mesenchymal stem cells from a large animal model. PLoS One, 9(11), e111533. https://doi.org/10.1371/journal.pone.0111533

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 9
Issue: 11
Pages: e111533
PII: e111533

Researcher Affiliations

MeSH Terms

  • Animals
  • Biomarkers / metabolism
  • Biphenyl Compounds / pharmacology
  • Cell Line
  • Cell Proliferation / drug effects
  • Gene Expression Regulation / drug effects
  • Horses
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Neurogenesis / drug effects
  • Neurons / cytology
  • Neurons / drug effects
  • Osteogenesis / drug effects
  • Phenethylamines / pharmacology
  • Receptors, Calcium-Sensing / agonists
  • Receptors, Calcium-Sensing / metabolism
  • Umbilical Cord / cytology

Conflict of Interest Statement

ERA-Amgen provided AMG641 for this study. (Amgen Inc., Thousand Oaks, California, United States of America; Research Program Agreement n ° 2011568566). This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

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Citations

This article has been cited 3 times.
  1. Wang T, Cao L, He S, Long K, Wang X, Yu H, Ma B, Xu X, Li W. Small RNA sequencing reveals a novel tsRNA-06018 playing an important role during adipogenic differentiation of hMSCs.. J Cell Mol Med 2020 Nov;24(21):12736-12749.
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  2. Wang T, Mei J, Li X, Xu X, Ma B, Li W. A novel tsRNA-16902 regulating the adipogenic differentiation of human bone marrow mesenchymal stem cells.. Stem Cell Res Ther 2020 Aug 24;11(1):365.
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  3. Sheehy EJ, Lemoine M, Clarke D, Gonzalez Vazquez A, O'Brien FJ. The Incorporation of Marine Coral Microparticles into Collagen-Based Scaffolds Promotes Osteogenesis of Human Mesenchymal Stromal Cells via Calcium Ion Signalling.. Mar Drugs 2020 Jan 23;18(2).
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