Improved isolation protocol for equine cord blood-derived mesenchymal stromal cells.
Abstract: A robust methodology for the isolation of cord blood-derived multipotent mesenchymal stromal cells (CB-MSCs) from fresh umbilical cord blood has not been reported in any species. The objective of this study was to improve the isolation procedure for equine CB-MSCs. Methods: Pre-culture separation of red and white blood cells was done using either PrepaCyte?-EQ medium or Ficoll-Paque? PREMIUM density medium. Regular FBS and MSC-qualified FBS were compared for their ability to support the establishment of putative primary MSC colonies. Conclusions: Our results indicate that PrepaCyte-EQ medium is superior to Ficoll-Paque PREMIUM density medium for the isolation of putative equine CB MSC and that MSC-qualified FBS may improve the isolation efficiency.
Publication Date: 2009-06-11 PubMed ID: 19513899DOI: 10.1080/14653240902887259Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses a study aiming to improve the isolation procedure for mesenchymal stromal cells (MSCs), specifically those derived from equine cord blood. The study compares different techniques for cell separation and media for primary MSC colony establishment.
Objective of the Research
- The main goal of the research was to better the methodology of isolation for equine cord blood-derived multipotent mesenchymal stromal cells (CB-MSCs). A well-established method of isolating these specific cells from fresh umbilical cord blood was missing, which made this research essential.
Methodology Employed
- To achieve this objective, the researchers carried out a pre-culture separation of red and white blood cells. This was done using two kinds of mediums: PrepaCyte-EQ and Ficoll-Paque PREMIUM density medium.
- The study also brought into comparison two types of Fetal Bovine Serum (FBS), Regular FBS and MSC-qualified FBS. These were examined for their ability to assist in the setup of putative primary MSC colonies.
Conclusions of the Study
- Based on the results of the study, the researchers concluded that the PrepaCyte-EQ medium is more efficient than Ficoll-Paque PREMIUM density medium for isolating equine CB-MSCs.
- Furthermore, they also found that the MSC-qualified FBS could potentially be more effective in improving the efficiency of the isolation process.
In sum, this research provides advancements in the isolation process of equine cord blood-derived mesenchymal stromal cells. Future studies could build on these findings to better the process further and to verify the efficiency of MSC-qualified FBS in the process.
Cite This Article
APA
Koch TG, Thomsen PD, Betts DH.
(2009).
Improved isolation protocol for equine cord blood-derived mesenchymal stromal cells.
Cytotherapy, 11(4), 443-447.
https://doi.org/10.1080/14653240902887259 Publication
Researcher Affiliations
- Department of Biomedical Sciences, University of Guelph, Ontario, Guelph, Canada. tkoch@uoguelph.ca
MeSH Terms
- Adipogenesis
- Animals
- Cell Separation / methods
- Chondrogenesis
- Colony-Forming Units Assay
- Fetal Blood / cytology
- Horses
- Leukocytes, Mononuclear / cytology
- Mesenchymal Stem Cells / cytology
- Osteogenesis
- Stromal Cells / cytology
Citations
This article has been cited 13 times.- Mayet A, Zablotski Y, Roth SP, Brehm W, Troillet A. Systematic review and meta-analysis of positive long-term effects after intra-articular administration of orthobiologic therapeutics in horses with naturally occurring osteoarthritis.. Front Vet Sci 2023;10:1125695.
- Mund SJK, MacPhee DJ, Campbell J, Honaramooz A, Wobeser B, Barber SM. Macroscopic, Histologic, and Immunomodulatory Response of Limb Wounds Following Intravenous Allogeneic Cord Blood-Derived Multipotent Mesenchymal Stromal Cell Therapy in Horses.. Cells 2021 Nov 1;10(11).
- Alizadeh Yegani A, Maytalman E, Kozanoglu I, Terzi MY, Aksu F. Biological behaviors of muscarinic receptors in mesenchymal stem cells derived from human placenta and bone marrow.. Iran J Basic Med Sci 2020 Jan;23(1):124-132.
- Mund SJK, Kawamura E, Awang-Junaidi AH, Campbell J, Wobeser B, MacPhee DJ, Honaramooz A, Barber S. Homing and Engraftment of Intravenously Administered Equine Cord Blood-Derived Multipotent Mesenchymal Stromal Cells to Surgically Created Cutaneous Wound in Horses: A Pilot Project.. Cells 2020 May 8;9(5).
- Roberts EL, Dang T, Lepage SIM, Alizadeh AH, Walsh T, Koch TG, Kallos MS. Improved expansion of equine cord blood derived mesenchymal stromal cells by using microcarriers in stirred suspension bioreactors.. J Biol Eng 2019;13:25.
- Sultana T, Lee S, Yoon HY, Lee JI. Current Status of Canine Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Veterinary Medicine.. Stem Cells Int 2018;2018:8329174.
- Desancé M, Contentin R, Bertoni L, Gomez-Leduc T, Branly T, Jacquet S, Betsch JM, Batho A, Legendre F, Audigié F, Galéra P, Demoor M. Chondrogenic Differentiation of Defined Equine Mesenchymal Stem Cells Derived from Umbilical Cord Blood for Use in Cartilage Repair Therapy.. Int J Mol Sci 2018 Feb 10;19(2).
- Williams LB, Co C, Koenig JB, Tse C, Lindsay E, Koch TG. Response to Intravenous Allogeneic Equine Cord Blood-Derived Mesenchymal Stromal Cells Administered from Chilled or Frozen State in Serum and Protein-Free Media.. Front Vet Sci 2016;3:56.
- Williams LB, Russell KA, Koenig JB, Koch TG. Aspiration, but not injection, decreases cultured equine mesenchymal stromal cell viability.. BMC Vet Res 2016 Mar 7;12:45.
- Tessier L, Bienzle D, Williams LB, Koch TG. Phenotypic and immunomodulatory properties of equine cord blood-derived mesenchymal stromal cells.. PLoS One 2015;10(4):e0122954.
- Williams LB, Tessier L, Koenig JB, Koch TG. Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro.. PLoS One 2014;9(12):e113615.
- Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
- Arufe MC, De la Fuente A, Fuentes I, Toro FJ, Blanco FJ. Umbilical cord as a mesenchymal stem cell source for treating joint pathologies.. World J Orthop 2011 Jun 18;2(6):43-50.
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