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Phenotypical and functional characteristics of mesenchymal stem cells derived from equine umbilical cord blood

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Abstract

Mesenchymal stem cells (MSCs) offer promise as therapeutic aid in the repair of tendon and ligament injuries in race horses. Fetal adnexa is considered as an ideal source of MSCs due to many advantages, including non-invasive nature of isolation procedures and availability of large tissue mass for harvesting the cells. However, MSCs isolated from equine fetal adnexa have not been fully characterized due to lack of species-specific markers. Therefore, this study was carried out to isolate MSCs from equine umbilical cord blood (UCB) and characterize them using cross-reactive markers. The plastic-adherent cells could be isolated from 13 out of 20 (65 %) UCB samples. The UCB derived cells proliferated till passage 20 with average cell doubling time of 46.40 ± 2.86 h. These cells expressed mesenchymal surface markers but did not express haematopoietic/leucocytic markers by RT-PCR and immunocytochemistry. The phenotypic expression of CD29, CD44, CD73 and CD90 was shown by 96.36 ± 1.28, 93.40 ± 0.70, 73.23 ± 1.29 and 46.75 ± 3.95 % cells, respectively in flow cytometry, whereas, reactivity against the haematopoietic antigens CD34 and CD45 was observed only in 2.4 ± 0.20 and 0.1 ± 0.0 % of cells, respectively. Osteogenic and chondrogenic differentiation could be achieved using established methods, whereas the optimum adipogenic differentiation was achieved after supplementing media with 15 % rabbit serum and 20 ng/ml of recombinant human insulin. In this study, we optimized methodology for isolation, cultural characterization, differentiation and immunophenotyping of MSCs from equine UCB. Protocols and markers used in this study can be employed for unequivocal characterization of equine MSCs.

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Acknowledgments

We thank National Research Centre on Equines (NRCE), Hisar for infrastructure support. Financial support to Niharika Mohanty from Indian Council of Agricultural Research (ICAR) and to Baldev R Gulati from Department of Biotechnology, Government of India is duly acknowledged. The authors wish to thank Equine Breeding Stud, Hisar, Haryana, India for providing access for sampling. The assistance in flow cytometry by BD-FACS academy, New Delhi, India is duly acknowledged.

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Authors and Affiliations

  1. Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, 125004, Haryana, India

    N. Mohanty, S. Gera & S. Kumar

  2. National Research Centre on Equines, Sirsa Road, Hisar, 125001, Haryana, India

    B. R. Gulati & R. Kumar

  3. Department of Veterinary Anatomy, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, 125004, Haryana, India

    P. Kumar

  4. Central Institute for Research on Buffaloes, Sirsa Road, Hisar, 125001, Haryana, India

    P. S. Yadav

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  1. N. Mohanty
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  2. B. R. Gulati
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  3. R. Kumar
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  4. S. Gera
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  5. S. Kumar
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  6. P. Kumar
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  7. P. S. Yadav
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Correspondence to B. R. Gulati.

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Mohanty, N., Gulati, B.R., Kumar, R. et al. Phenotypical and functional characteristics of mesenchymal stem cells derived from equine umbilical cord blood. Cytotechnology 68, 795–807 (2016). https://doi.org/10.1007/s10616-014-9831-z

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  • DOI: https://doi.org/10.1007/s10616-014-9831-z

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