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Frontiers in veterinary science2023; 10; 1154987; doi: 10.3389/fvets.2023.1154987

Influence of Rho/ROCK inhibitor Y-27632 on proliferation of equine mesenchymal stromal cells.

Abstract: Mesenchymal stromal cells (MSC) isolated form bone marrow and adipose tissue are the most common cells used for cell therapy of orthopedic diseases. MSC derived from different tissues show differences in terms of their proliferation, differentiation potential and viability in prolonged cell culture. This suggests that there may be subtle differences in intracellular signaling pathways that modulate these cellular characteristics. The Rho/ROCK signaling pathway is essential for many cellular functions. Targeting of this pathway by the ROCK inhibitor Y-27632 has been shown to be beneficial for cell viability and proliferation of different cell types. The aim of this study was to investigate the effects of Rho/ROCK inhibition on equine MSC proliferation using bone marrow-derived MSC (BMSC) and adipose-derived MSC (ASC). Primary ASC and BMSC were stimulated with or without 10 ng/mL TGF-β3 or 10 μM Y-27632, as well as both in combination. Etoposide at 10 μM was used as a positive control for inhibition of cell proliferation. After 48 h of stimulation, cell morphology, proliferation activity and gene expression of cell senescence markers p53 and p21 were assessed. ASC showed a trend for higher basal proliferation than BMSC, which was sustained following stimulation with TGF-β3. This included a higher proliferation with TGF-β3 stimulation compared to Y-27632 stimulation ( < 0.01), but not significantly different to the no treatment control when used in combination. Expression of p21 and p53 was not altered by stimulation with TGF-β3 and/or Y-27632 in either cell type. In summary, the Rho/ROCK inhibitor Y-27632 had no effect on proliferation activity and did not induce cell senescence in equine ASC and BMSC.
Copyright © 2023 Melzer, Burk, Guest and Dudhia.
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Publication Date: 2023-06-06 PubMed ID: 37346276PubMed Central: PMC10279950DOI: 10.3389/fvets.2023.1154987Google Scholar: Lookup
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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 study investigates the effect of Rho/ROCK inhibitor Y-27632 on the proliferation of mesenchymal stromal cells (MSC) derived from horses, specifically from their bone marrow and adipose tissue. The research discovered that the inhibitor had no significant effect on cell proliferation and did not induce cell senescence.

Objective of the Research

  • The core objective of this study was to understand the effects of Rho/ROCK inhibitor (Y-27632) on the proliferation of equine Mesenchymal Stromal Cells (MSCs) derived from bone marrow (BMSC) and adipose tissue (ASC).

Methodology

  • Both ASC and BMSC cells were stimulated with or without 10ng/mL TGF-β3 or 10μM Y-27632, individually and in combination.
  • 10μM etoposide was used as a positive control to influence the inhibition of cell proliferation.
  • Morphology of cells, their proliferation activity and gene expression of cell senescence markers p53 and p21 were assessed after 48 hours of stimulation.

Key Findings

  • ASC cells showed a slight trend for higher basal proliferation than BMSC cells, which was maintained even after stimulation with TGF-β3.
  • The research found higher proliferation in ASC cells with TGF-β3 compared to Y-27632. However, the difference was not significant when compared to the no treatment control when both TGF-β3 and Y-27632 were used in combination.
  • The expression of cell senescence markers p21 and p53 was not altered by stimulation with TGF-β3 and/or Y-27632 in either cell type signifying no induction of senescence.

Conclusion

  • In conclusion, it was found that the Rho/ROCK inhibitor, Y-27632 had no impact on cell proliferation and did not induce senescence in equine ASC and BMSC. This finding is crucial as it suggests that Y-27632 does not affect the viability or function of these cells, which are often used for therapeutic purposes.

Cite This Article

APA
Melzer M, Burk J, Guest DJ, Dudhia J. (2023). Influence of Rho/ROCK inhibitor Y-27632 on proliferation of equine mesenchymal stromal cells. Front Vet Sci, 10, 1154987. https://doi.org/10.3389/fvets.2023.1154987

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 10
Pages: 1154987
PII: 1154987

Researcher Affiliations

Melzer, Michaela
  • Equine Clinic (Surgery, Orthopedics), Faculty of Veterinary Medicine, Justus Liebig University, Giessen, Germany.
Burk, Janina
  • Equine Clinic (Surgery, Orthopedics), Faculty of Veterinary Medicine, Justus Liebig University, Giessen, Germany.
Guest, Deborah J
  • Department of Clinical Sciences and Services, Royal Veterinary College, Hertfordshire, United Kingdom.
Dudhia, Jayesh
  • Department of Clinical Sciences and Services, Royal Veterinary College, Hertfordshire, United Kingdom.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Citations

This article has been cited 1 times.
  1. Shi J, Cai J, Kong L, Ying L, Liu X, Jiang M, Pan D. Upregulation of miR-183 inhibits the invasion and migration of endometrial stromal cells in endometriosis patients by downregulating Ezrin. Front Oncol 2025;15:1537528.
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