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Home / Equine Research Bank / Front Vet Sci / Evaluation of stability and safety of equine mesenchymal ste...
Frontiers in veterinary science2024; 11; 1330009; doi: 10.3389/fvets.2024.1330009

Evaluation of stability and safety of equine mesenchymal stem cells derived from amniotic fluid for clinical application.

Abstract: Amniotic fluid mesenchymal stem cells (AF-MSCs), which can be obtained from fetal tissue, reportedly have self-renewal capacity and multi-lineage differentiation potential. The aim of this study was to identify the biological characteristics of AF-MSCs and evaluate their stability and safety in long-term culture. To confirm the biological characteristics of AF-MSCs, morphology, proliferation capacity, karyotype, differentiation capacity, gene expression level, and immunophenotype were analyzed after isolating AF-MSCs from equine amniotic fluid. AF-MSCs were differentiated into adipocytes, chondrocytes, and osteocytes. Immunophenotype analyses revealed expression levels of ≥95% and ≤ 2% of cells for a positive and negative marker, respectively. Analysis of the MSCs relative gene expression levels of AF-MSCs was approximately at least twice that of the control. The endotoxin level was measured to verify the safety of AF-MSCs and was found to be less than the standard value of 0.5 EU/ml. AF-MSCs were cultured for a long time without any evidence of abnormalities in morphology, proliferation ability, and karyotype. These results suggest that amniotic fluid is a competent source for acquiring equine MSCs and that it is valuable as a cell therapy due to its high stability.
Copyright © 2024 Kim, Lee, Kim, Kil and Kim.
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Publication Date: 2024-02-14 PubMed ID: 38420210PubMed Central: PMC10899390DOI: 10.3389/fvets.2024.1330009Google 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.

Overview

  • The study investigates the characteristics, stability, and safety of mesenchymal stem cells derived from equine amniotic fluid (AF-MSCs) to assess their suitability for clinical applications.

Introduction to AF-MSCs

  • Amniotic fluid mesenchymal stem cells (AF-MSCs) are stem cells isolated from the amniotic fluid surrounding a fetus.
  • These stem cells are known for their ability to self-renew and differentiate into multiple types of cells (multi-lineage differentiation), making them promising for regenerative medicine and cell therapy.
  • In horses, AF-MSCs could be valuable for treating injuries or diseases, but their biological properties and safety need to be thoroughly validated.

Objectives of the Study

  • To isolate mesenchymal stem cells from equine amniotic fluid.
  • To characterize the biological features of these AF-MSCs in terms of morphology, growth, genetic stability, differentiation capability, gene expression, and immune markers.
  • To assess the stability and safety of AF-MSCs during extended culture periods, especially evaluating for potential abnormalities.

Methods and Analyses

  • Isolation and Morphology: Stem cells were successfully isolated from equine amniotic fluid, and their cell shape and structure (morphology) were observed for normal features.
  • Proliferation Capacity: The growth rate and ability of AF-MSCs to multiply were measured over time to ensure robust expansion.
  • Karyotyping: Chromosomal analysis was performed to check for genetic stability and detect any abnormalities.
  • Differentiation Capacity: The ability of AF-MSCs to differentiate into three primary cell types—adipocytes (fat cells), chondrocytes (cartilage cells), and osteocytes (bone cells)—was tested, confirming their multi-lineage potential.
  • Gene Expression Analysis: The expression levels of genes related to stemness and mesenchymal cell identity were assessed, showing approximately double expression compared to control cells, indicating strong stem cell characteristics.
  • Immunophenotyping: Cell surface markers were analyzed to confirm mesenchymal stem cell identity; ≥95% cells expressed positive MSC markers, and ≤2% expressed negative markers.
  • Endotoxin Level Measurement: To ensure safety, endotoxin contamination was evaluated; levels were below the acceptable threshold (0.5 EU/ml), indicating no harmful contamination.
  • Long-term Culture Evaluation: AF-MSCs were cultured for an extended period and showed no abnormalities in morphology, proliferation, or karyotype, suggesting stability over time.

Key Results

  • AF-MSCs showed typical mesenchymal stem cell morphology and maintained healthy proliferation rates.
  • Chromosome analysis (karyotype) demonstrated genetic stability with no abnormalities after prolonged culture.
  • Successful differentiation into adipocytes, chondrocytes, and osteocytes confirmed the multi-lineage differentiation potential of AF-MSCs.
  • Gene expression of MSC-related markers was significantly higher compared to control cells, highlighting robust stem cell properties.
  • Immunophenotype analysis confirmed that the vast majority of cells carried expected surface markers typical of MSCs.
  • Endotoxin levels were safely below concerning thresholds, reducing the risk of adverse immune or inflammatory reactions in clinical use.
  • No detrimental changes were observed during long-term culture, supporting the cells’ stability for therapeutic applications.

Conclusions and Implications

  • Equine amniotic fluid is a reliable and effective source of mesenchymal stem cells suitable for clinical and therapeutic use.
  • The isolated AF-MSCs exhibit high proliferative ability, multi-lineage differentiation capacity, and genetic stability, crucial for safe and effective cell therapies.
  • The safety evaluation, including endotoxin testing and long-term culture monitoring, supports the practical use of these cells without significant risk of genetic or functional abnormalities.
  • This research underpins the potential of equine AF-MSCs as a valuable tool in regenerative veterinary medicine, particularly for treatments requiring stable and safe stem cell populations.

Cite This Article

APA
Kim EY, Lee EJ, Kim RE, Kil TY, Kim MK. (2024). Evaluation of stability and safety of equine mesenchymal stem cells derived from amniotic fluid for clinical application. Front Vet Sci, 11, 1330009. https://doi.org/10.3389/fvets.2024.1330009

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 11
Pages: 1330009
PII: 1330009

Researcher Affiliations

Kim, Eun Young
  • MK Biotech Co., Ltd., Daejeon, Republic of Korea.
Lee, Eun Ji
  • Division of Animal and Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
Kim, Ryoung Eun
  • MK Biotech Co., Ltd., Daejeon, Republic of Korea.
Kil, Tae Young
  • Department of Social Welfare, Joongbu University, Chungcheongnam-do, Republic of Korea.
Kim, Min Kyu
  • MK Biotech Co., Ltd., Daejeon, Republic of Korea.
  • Division of Animal and Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.

Conflict of Interest Statement

EK, RK, and MK are employed by MK Biotech Co., Ltd. The remaining 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 2 times.
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  2. Shi L, Khan MZ, Ullah A, Liang H, Geng M, Akhtar MF, Na J, Han Y, Wang C. Advancements in Stem Cell Applications for Livestock Research: A Review.. Vet Sci 2025 Apr 23;12(5).
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