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Home / Equine Research Bank / BMC Vet Res / Culture of mesenchymal stem cells derived from equine synovi...
BMC veterinary research2018; 14(1); 114; doi: 10.1186/s12917-018-1425-0

Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules.

Abstract: Mesenchymal stem cells derived from the synovial membrane (MSCSM) have a greater potential for joint regeneration, besides the capacity for chondrogenic differentiation, since they are a source closer to the chondrocytes. This study aimed to cultivate and evaluate viability and differentiation of MSC encapsulated in a three-dimensional alginate hydrogel (HA) scaffold. Samples of the synovial membrane of the metatarsophalangeal joint of 4 horses were collected by astroscopic surgery. These were subjected to enzymatic digestion, isolated mesenchymal cells, cultured in monolayers and encapsulated at various concentrations, 10; 20; 50; 10; 20 cells in 1.5% sodium alginate solution. The gelatinization process was carried out and cultured for 4 weeks. Viability and cell proliferation were performed by dissolving the microcapsules and counting with trypan blue. The ratio of live cells and total live cells at intervals 0, 7, 14, 21 and 28 days was analyzed. For the evaluation of differentiation, histological sections stained with hematoxylin and eosin and toluidine blue were performed. There was no statistical difference in the proportion of live cells between groups over the 28 days. The group of 10 cells obtained a higher total number of living cells at the end of the experiment. Through the histological analysis it was possible to observe at 7 days a low amount of spherical cells with chondrocyte characteristics. On day 21, chondrogenic differentiation became evident, with pericellular and territorial matrix production. This study demonstrated the efficiency of HA as a scaffold for MSC and the chondrogenic differentiation, promising for use in the treatment of joint injuries in horses.
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Publication Date: 2018-03-27 PubMed ID: 29587733PubMed Central: PMC5870504DOI: 10.1186/s12917-018-1425-0Google 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.

The research examines the potential of using equine mesenchymal stem cells derived from the synovial membrane, encased in an alginate hydrogel scaffold, for joint regeneration. The study assessed the cell viability and differentiation over four weeks, concluding that the method is efficient and holds promise for treating joint injuries in horses.

Methods

  • The source of the mesenchymal stem cells for the research were the synovial membranes of the metatarsophalangeal joint of four horses, acquired through arthroscopic surgery.
  • The membranes were subjected to enzymatic digestion, which broke down the component tissues and allowed for the isolation of the mesenchymal cells.
  • The isolated cells were grown in monolayers. This is a culture of cells grown in a thin, consistent layer; typically, this is conducive to cell proliferation and maintains cells in a healthy state.
  • The cells were encapsulated in a 1.5% sodium alginate solution at five different concentrations (10, 20, 50, 10, 20 cells).
  • The encapsulation process involved gelatinization and culturing for four weeks.

Assessment of Viability and Proliferation

  • To assess the viability and proliferation of the cells, the microcapsules were dissolved and the cells counted using trypan blue, a dye that can distinguish live from dead cells.
  • This assessment took place at several intervals: on days 0, 7, 14, 21 and 28.

Assessment of Differentiation

  • To analyze the differentiation of the mesenchymal stem cells, histological samples of the cultured cells were taken.
  • These samples were stained with hematoxylin and eosin as well as toluidine blue, dyes that highlight the microscopic structure and morphology of cells.
  • Chondrocyte characteristics (cells that form cartilage) were evident in some of the cells as early as day 7.
  • By day 21, chondrogenic differentiation, the process by which cells become specialized chondrocyte cells, was clear, evidenced by the generation of pericellular and territorial matrix, types of extracellular matrix materials essential for cartilage function.

Outcome

  • No significant difference was found in the proportion of live cells between the groups on any of the 28 days.
  • The 10-cell group displayed the highest total number of live cells at the end of the experiment, suggesting the suitability of the method at low cell concentrations.
  • The histological analyses confirmed the successful chondrogenic differentiation of the mesenchymal stem cells.
  • Therefore, the alginate hydrogel scaffold proved an effective medium for culturing these mesenchymal stem cells and promoting their differentiation, supporting the potential for using this technique in the treatment of horse joint injuries.

Cite This Article

APA
Santos VH, Pfeifer JPH, de Souza JB, Milani BHG, de Oliveira RA, Assis MG, Deffune E, Moroz A, Alves ALG. (2018). Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules. BMC Vet Res, 14(1), 114. https://doi.org/10.1186/s12917-018-1425-0

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 14
Issue: 1
Pages: 114
PII: 114

Researcher Affiliations

Santos, Vitor Hugo
  • Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.
Pfeifer, João Pedro Hübbe
  • Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.
de Souza, Jaqueline Brandão
  • Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.
Milani, Betsabéia Heloisa Gentilha
  • Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.
de Oliveira, Rogério Antonio
  • Departament of Statistics, Institute of Biosciences, UNESP, District of Rubião Júnior, s / n, Botucatu, SP, Brazil.
Assis, Marjorie Golim
  • Departament of Graduate Program in Research and Development: Medical Biotechnology (Professional Master's) from the Blood Center of UNESP, Blood Centre Division, District of Rubião Júnior, s / n° -, Botucatu, SP, Brazil.
Deffune, Elenice
  • Departament of Urology, University of Medicine, UNESP, District of Rubião Junior s / n° - Blood Centre Division - Laboratory of Cellular Engineering, Botucatu, SP, Brazil.
Moroz, Andrei
  • Departament of Bioprocesses and Biotechnology, FCFAR - UNESP, Rodovia Araraquara Jaú, KM 01, São Paulo, Brazil.
Alves, Ana Liz Garcia
  • Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil. anaalves@fmvz.unesp.br.

MeSH Terms

  • Alginates
  • Animals
  • Capsules
  • Cell Culture Techniques / veterinary
  • Cell Survival
  • Female
  • Glucuronic Acid
  • Hexuronic Acids
  • Horses
  • Hydrogels
  • Male
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / physiology

Conflict of Interest Statement

ETHICS APPROVAL AND CONSENT TO PARTICIPATE: The experiment was carried out at the Veterinary Hospital of the University of Veterinary Medicine and Zootechny (FMVZ), UNESP, Campus de Botucatu (Department of Surgery and Veterinary Anesthesiology), in the Laboratory of Cell Therapy, FMVZ, Unesp Campus Botucatu) in 2016. The owner of the animals signed the consent form to use their animals in the study. It was approved by the Ethics Committee on Animal Experimentation of the University of Veterinary Medicine and Animal Science of the State University of São Paulo (UNESP) - Botucatu, under protocol number 052/2016. CONSENT FOR PUBLICATION: Not applicable COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER’S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Citations

This article has been cited 7 times.
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