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Home/Equine Research Bank/BMC Biotechnol/Isolation of mesenchymal stem cells from equine umbilical co...
BMC biotechnology2007; 7; 26; doi: 10.1186/1472-6750-7-26

Isolation of mesenchymal stem cells from equine umbilical cord blood.

Abstract: There are no published studies on stem cells from equine cord blood although commercial storage of equine cord blood for future autologous stem cell transplantations is available. Mesenchymal stem cells (MSC) have been isolated from fresh umbilical cord blood of humans collected non-invasively at the time of birth and from sheep cord blood collected invasively by a surgical intrauterine approach. Mesenchymal stem cells isolation percentage from frozen-thawed human cord blood is low and the future isolation percentage of MSCs from cryopreserved equine cord blood is therefore expectedly low. The hypothesis of this study was that equine MSCs could be isolated from fresh whole equine cord blood. Results: Cord blood was collected from 7 foals immediately after foaling. The mononuclear cell fraction was isolated by Ficoll density centrifugation and cultured in a DMEM low glucose based media at 38.5 degrees C in humidified atmosphere containing 5% CO2. In 4 out of 7 samples colonies with MSC morphology were observed. Cellular morphology varied between monolayers of elongated spindle-shaped cells to layered cell clusters of cuboidal cells with shorter cytoplasmic extensions. Positive Alizarin Red and von Kossa staining as well as significant calcium deposition and alkaline phosphatase activity confirmed osteogenesis. Histology and positive Safranin O staining of matrix glycosaminoglycans illustrated chondrogenesis. Oil Red O staining of lipid droplets confirmed adipogenesis. Conclusions: We here report, for the first time, the isolation of mesenchymal-like stem cells from fresh equine cord blood and their differentiation into osteocytes, chondrocytes and adipocytes. This novel isolation of equine cord blood MSCs and their preliminary in vitro differentiation positions the horse as the ideal pre-clinical animal model for proof-of-principle studies of cord blood derived MSCs.
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Publication Date: 2007-05-30 PubMed ID: 17537254PubMed Central: PMC1904213DOI: 10.1186/1472-6750-7-26Google 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 demonstrates for the first time that mesenchymal-like stem cells can be isolated from fresh equine cord blood, differentiating into osteocytes, chondrocytes and adipocytes. This suggests the horse as an ideal pre-clinical animal model for proof-of principle studies of cord blood derived mesenchymal stem cells.

Research Objectives and Hypothesis

  • The researchers aimed to isolate mesenchymal stem cells (MSC) from fresh equine umbilical cord blood, which had not been studied or documented before.
  • The study hypothesized that MSCs, already known to be isolable from human and sheep cord blood, could also be isolated from equine cord blood.

Methodology

  • Cord blood was collected from 7 foals immediately after their birth.
  • The mononuclear cell fraction was then isolated by Ficoll density centrifugation and cultured in a specially created environment (DMEM low glucose based media at 38.5 degrees C in humidified atmosphere containing 5% CO2).
  • Using this method, MSC-like colonies were observed in 4 out of 7 samples. The cellular morphology in these colonies varied widely.

Results and Observations

  • Cultures displayed positive Alizarin Red and von Kossa staining, indicating calcium deposition and alkaline phosphatase activity, thus confirming osteogenesis (formation of bone cells).
  • Histology and positive Safranin O staining of matrix glycosaminoglycans (complex carbohydrates) demonstrated chondrogenesis (formation of cartilage cells).
  • Oil Red O staining of lipid droplets illustrated adipogenesis (formation of fat cells).

Conclusions and Implications

  • The study successfully isolated mesenchymal-like stem cells from fresh equine cord blood and induced them to differentiate into osteocytes, chondrocytes and adipocytes.
  • This novel discovery holds significance as it supports the horse as an ideal pre-clinical animal model for proof-of-principle studies of cord blood-derived MSCs.
  • This research could pave the way for future studies and prospective applications related to equine cord blood MSCs, even though the isolation percentage from frozen-thawed cord blood is expected to be low.

Cite This Article

APA
Koch TG, Heerkens T, Thomsen PD, Betts DH. (2007). Isolation of mesenchymal stem cells from equine umbilical cord blood. BMC Biotechnol, 7, 26. https://doi.org/10.1186/1472-6750-7-26

Publication

BMC biotechnology
ISSN: 1472-6750
NlmUniqueID: 101088663
Country: England
Language: English
Volume: 7
Pages: 26

Researcher Affiliations

Koch, Thomas G
  • Department of Biomedical Sciences, University of Guelph, Guelph, ON, Canada. tkoch@uoguelph.ca
Heerkens, Tammy
    Thomsen, Preben D
      Betts, Dean H

        MeSH Terms

        • Adipocytes / cytology
        • Animals
        • Animals, Newborn
        • Cell Culture Techniques / methods
        • Cell Differentiation
        • Cell Separation / methods
        • Cells, Cultured
        • Chondrocytes / cytology
        • Cryopreservation / methods
        • Fetal Blood / cytology
        • Horses
        • Mesenchymal Stem Cells / cytology
        • Osteocytes / cytology
        • Tissue Engineering / methods

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