Isolation and characterization of equine amniotic fluid-derived multipotent stem cells.
Abstract: Amniotic fluid (AF) is a well-known source of stem cells. However, there have been no reports regarding equine AF stem cells. We have isolated equine AF-derived multipotent stem cells (MSC) (eAF-MSC) and show that these cells exhibit self-renewal ability and multilineage differentiation. Methods: AF was obtained from thoroughbred mares and mononuclear cells (MNC) were isolated by Ficoll-Paque density gradient. We measured the cumulative population doubling level (CPDL) and characterized the immunophenotype by flow cytometry. To investigate differentiation ability, a trilineage differentiation assay was conducted. Results: eAF-MSC could be isolated and the proliferation level was high. eAF-MSC presented typical MSC phenotypic markers, as determined by flow cytometry. Moreover, eAF-MSC showed a trilineage differentiation capability. Conclusions: Equine AF is a good source of MSC. Furthermore, eAF-MSC may be useful as a cell therapy application for horses.
Publication Date: 2010-09-22 PubMed ID: 20860427DOI: 10.3109/14653249.2010.520312Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the potential of equine amniotic fluid as a source of multipotent stem cells, demonstrating their ability for self-renewal and differentiation into multiple cell types which could have applications in horse cell therapy.
Methods
- The research team collected amniotic fluid from thoroughbred mares. The fluid was then processed to isolate mononuclear cells (MNC), a type of stem cell, using Ficoll-Paque density gradient, a common technique used to separate different types of cells based on their density.
- Next, they tracked the cumulative population doubling level (CPDL), which is a measurement of the cells’ ability to proliferate, or multiply. High CPDL signifies a high level of neural stem cell proliferation.
- The team then characterized the cells’ immunophenotype using flow cytometry, a technique that allows identification and counting of different types of cells in a sample. This confirmed the presence of typical markers associated with multipotent stem cells (MSC), essentially determining the identity of the cells.
- The researchers conducted a trilineage differentiation assay to study the cells’ differentiation capacity. This is a lab process involving the application of certain stimulants to the cells to spur them to change, or differentiate, into other cell types—in this case testing the MSC’s ability to become three different cell types.
Results
- The results showed successful isolation of eAF-MSC (Equine Amniotic Fluid-Derived Multipotent Stem Cells), which demonstrated a high proliferation level – this suggests that they can reproduce and increase in number effectively.
- The cells presented typical MSC phenotypic markers when examined by flow cytometry, confirming that these cells were indeed multipotent stem cells.
- Furthermore, the cells exhibited trilineage differentiation capability, meaning they were not only able to reproduce, but could also differentiate into three different cell types.
Conclusions
- This research concluded that equine amniotic fluid is a viable source of multipotent stem cells.
- The multipotent stem cells derived from this fluid exhibited both high proliferation capacity and the ability to differentiate into multiple cell types.
- This points to the potential to utilise these cells in cell therapy applications for horses, opening up possibilities for new treatments or therapies in equine medicine.
Cite This Article
APA
Park SB, Seo MS, Kang JG, Chae JS, Kang KS.
(2010).
Isolation and characterization of equine amniotic fluid-derived multipotent stem cells.
Cytotherapy, 13(3), 341-349.
https://doi.org/10.3109/14653249.2010.520312 Publication
Researcher Affiliations
- Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
MeSH Terms
- Adipogenesis
- Amniotic Fluid / cytology
- Animals
- Cell Lineage
- Cell Proliferation
- Cell Separation / methods
- Cells, Cultured
- Chondrogenesis
- Flow Cytometry
- Horses
- Humans
- Immunophenotyping
- Multipotent Stem Cells / cytology
- Multipotent Stem Cells / metabolism
- Osteogenesis
- Sequence Homology, Amino Acid
Citations
This article has been cited 14 times.- Park A, Oh HJ, Ji K, Choi EM, Kim D, Kim E, Kim MK. Effect of Passage Number of Conditioned Medium Collected from Equine Amniotic Fluid Mesenchymal Stem Cells: Porcine Oocyte Maturation and Embryo Development. Int J Mol Sci 2022 Jun 12;23(12).
- Sogawa K, Okawa R, Yachiku K, Shiozaki M, Miura T, Takayanagi H, Shibata T, Ezoe S. Effects of continuous exposure to low concentration of ClO(2) gas on the growth, viability, and maintenance of undifferentiated MSCs in long-term cultures. Regen Ther 2020 Jun;14:184-190.
- Barboni B, Russo V, Berardinelli P, Mauro A, Valbonetti L, Sanyal H, Canciello A, Greco L, Muttini A, Gatta V, Stuppia L, Mattioli M. Placental Stem Cells from Domestic Animals: Translational Potential and Clinical Relevance. Cell Transplant 2018 Jan;27(1):93-116.
- Kannaiyan J, Muthukutty P, Iqbal MDT, Paulraj B. Villous Chorion: A Potential Source for Pluripotent-like Stromal Cells. J Nat Sci Biol Med 2017 Jul-Dec;8(2):221-228.
- Da Sacco S, Perin L, Sedrakyan S. Amniotic fluid cells: current progress and emerging challenges in renal regeneration. Pediatr Nephrol 2018 Jun;33(6):935-945.
- Rohban R, Pieber TR. Mesenchymal Stem and Progenitor Cells in Regeneration: Tissue Specificity and Regenerative Potential. Stem Cells Int 2017;2017:5173732.
- Lo Surdo JL, Millis BA, Bauer SR. Automated microscopy as a quantitative method to measure differences in adipogenic differentiation in preparations of human mesenchymal stromal cells. Cytotherapy 2013 Dec;15(12):1527-40.
- Kang JG, Park SB, Seo MS, Kim HS, Chae JS, Kang KS. Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood. J Vet Sci 2013;14(3):367-71.
- Seo MS, Park SB, Kim HS, Kang JG, Chae JS, Kang KS. Isolation and characterization of equine amniotic membrane-derived mesenchymal stem cells. J Vet Sci 2013;14(2):151-9.
- Choi SA, Choi HS, Kim KJ, Lee DS, Lee JH, Park JY, Kim EY, Li X, Oh HY, Lee DS, Kim MK. Isolation of canine mesenchymal stem cells from amniotic fluid and differentiation into hepatocyte-like cells. In Vitro Cell Dev Biol Anim 2013 Jan;49(1):42-51.
- Spaas JH, Guest DJ, Van de Walle GR. Tendon regeneration in human and equine athletes: Ubi Sumus-Quo Vadimus (where are we and where are we going to)?. Sports Med 2012 Oct 1;42(10):871-90.
- Lo Surdo J, Bauer SR. Quantitative approaches to detect donor and passage differences in adipogenic potential and clonogenicity in human bone marrow-derived mesenchymal stem cells. Tissue Eng Part C Methods 2012 Nov;18(11):877-89.
- Iacono E, Cunto M, Zambelli D, Ricci F, Tazzari PL, Merlo B. Could fetal fluid and membranes be an alternative source for mesenchymal stem cells (MSCs) in the feline species? A preliminary study. Vet Res Commun 2012 Jun;36(2):107-18.
- Lovati AB, Corradetti B, Lange Consiglio A, Recordati C, Bonacina E, Bizzaro D, Cremonesi F. Comparison of equine bone marrow-, umbilical cord matrix and amniotic fluid-derived progenitor cells. Vet Res Commun 2011 Feb;35(2):103-21.
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