Expression of reprogramming factors in mesenchymal stem cells isolated from equine umbilical cord Wharton’s jelly and amniotic fluid

N S RATHORE; S K KASHYAP; ANUPAMA DEORA; PANKAJ KUMAR; J SINGH; T R TALLURI

doi:10.56093/ijans.v91i2.113818

Authors

N S RATHORE ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
S K KASHYAP ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
ANUPAMA DEORA ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
PANKAJ KUMAR ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
J SINGH ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
T R TALLURI ICAR- National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India

https://doi.org/10.56093/ijans.v91i2.113818

Keywords:

Equine, Mesenchymal stem cells, Reprogramming factors, Umbilical cord, Wharton’s jelly

Abstract

Stem cells represent the most promising population for regenerative cell therapy and have gained much attention during the recent past. Mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into trilineages. Like haematopoietic cells, mesenchymal cells have been shown to proliferate and form fibroblast-like colonies in vitro. Despite major progress in our general knowledge related to the application of adult stem cells, finding alternative sources for bone marrow MSCs has remained a challenge. A wide diversity of isolation procedures for mesenchymal stromal cells from various tissues of the placenta, umbilical cord and Wharton's jelly have been described for humans and other species. In this study, we isolated established umbilical cord Whartonâ€™s jelly as a primary source for isolation of mesenchymal stem cells since it is a rich source of stem cells and no ethical concerns are involved. Equine umbilical cord Whartonâ€™s jelly segments were collected during foaling time and digested enzymatically and cultured in-vitro in culture medium. In addition to the study of their morphology and colony forming units, the expression of reprogramming factors by the isolated MSCs were also studied. The isolated MSCs were observed to be plastic adherent, clonogenic and their morphology were polygonal, star shaped and fibroblast like. They revealed a strong expression of pluripotent stemness markers OCT-4, SOX-2, Nanog and KLF-4. From the current study, it can be concluded that Wharton's jelly is a rich source of stem cells with stemness properties expressing the reprogramming factors and mesenchymal like morphology and could be used as an alternate for the bone marrow derived mesenchymal stem cells for cell based regenerative therapies.

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References

Barry F P and Murphy J M. 2004. Mesenchymal stem cells: clinical applications and biological characterization. International Journal of Biochemistry and Cell Biology 36(4): 568–84. DOI: https://doi.org/10.1016/j.biocel.2003.11.001

Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S and Smith A. 2003. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113(5): 643–55.

Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S and Smith A. 2003. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113(5): 643–55. DOI: https://doi.org/10.1016/S0092-8674(03)00392-1

Cremonesi F, Violini S, Consiglio A L, Ramelli P, Ranzenigo G and Mariani P. 2008. Isolation, in vitro culture and characterization of foal umbilical cord stem cells at birth. Veterinary Research Communications 32(1): 139–42. DOI: https://doi.org/10.1007/s11259-008-9116-0

De Coppi P, Bartsch G, Siddiqui M M, Xu T, Santos C C, Perin L, Mostoslavsky G, Serre A C, Snyder E Y and Yoo J J. 2007. Isolation of amniotic stem cell lines with potential for therapy. Nature Biotechnology 25(1): 100–06. DOI: https://doi.org/10.1038/nbt1274

D’ippolito G, Schiller P C, Ricordi C, Roos B A and Howard G A. 1999. Age related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. Journal of Bone and Mineral Research 14(7): 1115–22. DOI: https://doi.org/10.1359/jbmr.1999.14.7.1115

Guest D J, Ousey J C and Smith M R. 2008. Defining the expression of marker genes in equine mesenchymal stromal cells. Stem Cells and Cloning: Advances and Applications 1: 1. DOI: https://doi.org/10.2147/SCCAA.S3824

Gulati B R, Kumar R, Mohanty N, Gera S, Kumar P and Yadav P S. 2016. Comparative evaluation of equine mesenchymal stem cells derived from amniotic fluid and umbilical cord blood. Indian Journal of Animal Sciences 86(5): 539–44.

Gulati B R, Kumar R, Mohanty N, Kumar P, Somasundaram R K and Yadav P S. 2013. Bone morphogenetic protein-12 induces tenogenic differentiation of mesenchymal stem cells derived from equine amniotic fluid. Cells Tissues Organs 198(5): 377– 89. DOI: https://doi.org/10.1159/000358231

Iacono E, Brunori L, Pirrone A, Pagliaro P P, Ricci F, Tazzari P L and Merlo B. 2012. Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton’s jelly in the horse. Reproduction 143(4): 455. DOI: https://doi.org/10.1530/REP-10-0408

Karahuseyinoglu S, Cinar O, Kilic E, Kara F, Akay G G, Demiralp D Ö, Tukun A, Uckan D and Can A. 2007. Biology of stem cells in human umbilical cord stroma: in situ and in vitro surveys. Stem cells 25(2): 319–31. DOI: https://doi.org/10.1634/stemcells.2006-0286

Kasashima Y, Ueno T, Tomita A, Goodship A and Smith R. 2011. Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells. Equine Veterinary Journal 43(3): 288–94. DOI: https://doi.org/10.1111/j.2042-3306.2010.00215.x

Kisiday J D, Kopesky P W, Evans C H, Grodzinsky A J, McIlwraith C W and Frisbie D D. 2008. Evaluation of adult equine bone marrow and adipose derived progenitor cell chondrogenesis in hydrogel cultures. Journal of Orthopaedic Research 26(3): 322–31. DOI: https://doi.org/10.1002/jor.20508

Koch T G, Heerkens T, Thomsen P D and Betts D H. 2007. Isolation of mesenchymal stem cells from equine umbilical cord blood. BMC Biotechnology 7(1): 1–9. DOI: https://doi.org/10.1186/1472-6750-7-26

Koerner J, Nesic D, Romero J D, Brehm W, Mainil Varlet P and Grogan S P. 2006. Equine peripheral blood derived progenitors in comparison to bone marrow derived mesenchymal stem cells. Stem Cells 24(6): 1613–19. DOI: https://doi.org/10.1634/stemcells.2005-0264

Lee K B, Hui J H, Song I C, Ardany L and Lee E H. 2007. Injectable mesenchymal stem cell therapy for large cartilage defects—a porcine model. Stem Cells 25(11): 2964–71. DOI: https://doi.org/10.1634/stemcells.2006-0311

Lovati A B, Corradetti B, Consiglio A L, Recordati C, Bonacina E, Bizzaro D and Cremonesi F. 2011. Comparison of equine bone marrow-, umbilical cord matrix and amniotic fluid- derived progenitor cells. Veterinary Research Communications 35(2): 103–21. DOI: https://doi.org/10.1007/s11259-010-9457-3

Pappa K I and Anagnou N P. 2009. Novel sources of fetal stem cells: Where do they fit on the developmental continuum? Regenerative Medicine 4(3): 423–33. DOI: https://doi.org/10.2217/rme.09.12

Park C Y, Kim D H, Son J S, Sung J J, Lee J, Bae S, Kim J H, Kim D W and Kim J S. 2015. Functional correction of large factor VIII gene chromosomal inversions in hemophilia A patient-derived iPSCs using CRISPR-Cas9. Cell Stem Cell 17(2): 213–20. DOI: https://doi.org/10.1016/j.stem.2015.07.001

Park S B, Seo M S, Kang J G, Chae J S and Kang K S. 2011. Isolation and characterization of equine amniotic fluid-derived multipotent stem cells. Cytotherapy 13(3): 341–49. DOI: https://doi.org/10.3109/14653249.2010.520312

Rathore N, Kashyap S, Deora A, Kumar P, Singh J, Tripathi B and Talluri T. 2018. Isolation and culture of putative mesenchymal stem cells from the equine amniotic membrane. Veterinary Practitioner 19(1): 180–83.

Reed S A and Johnson S E. 2008. Equine umbilical cord blood contains a population of stem cells that express Oct4 and differentiate into mesodermal and endodermal cell types. Journal of Cellular Physiology 215(2): 329–36. DOI: https://doi.org/10.1002/jcp.21312

Schuh E M, Friedman M S, Carrade D D, Li J, Heeke D, Oyserman S M, Galuppo L D, Lara D J, Walker N J and Ferraro G L. 2009. Identification of variables that optimize isolation and culture of multipotent mesenchymal stem cells from equine umbilical-cord blood. American Journal of Veterinary Research 70(12): 1526–35. DOI: https://doi.org/10.2460/ajvr.70.12.1526

Slaymaker I M, Gao L, Zetsche B, Scott D A, Yan W X and Zhang F. 2016. Rationally engineered Cas9 nucleases with improved specificity. Science 351(6268): 84–88. DOI: https://doi.org/10.1126/science.aad5227

Talluri T R, Kumar D, Glage S, Garrels W, Ivics Z, Debowski K, Behr R and Kues W A. 2014. Non-viral reprogramming of fibroblasts into induced pluripotent stem cells by Sleeping Beauty and piggyBac transposons. Biochemical and Biophysical Research Communications 450(1): 581–87. DOI: https://doi.org/10.1016/j.bbrc.2014.06.014

Toupadakis C A, Wong A, Genetos D C, Cheung W K, Borjesson D L, Ferraro G L, Galuppo L D, Leach J K, Owens S D and Yellowley C E. 2010. Comparison of the osteogenic potential of equine mesenchymal stem cells from bone marrow, adipose tissue, umbilical cord blood, and umbilical cord tissue. American Journal of Veterinary Research 71(10): 1237–45. DOI: https://doi.org/10.2460/ajvr.71.10.1237

Vidal M A, Kilroy G E, Johnson J R, Lopez M J, Moore R M and Gimble J M. 2006. Cell growth characteristics and differentiation frequency of adherent equine bone marrow– derived mesenchymal stromal cells: Adipogenic and osteogenic capacity. Veterinary Surgery 35(7): 601–10. DOI: https://doi.org/10.1111/j.1532-950X.2006.00197.x

Violini S, Ramelli P, Pisani L F, Gorni C and Mariani P. 2009. Horse bone marrow mesenchymal stem cells express embryo stem cell markers and show the ability for tenogenic differentiation by in vitro exposure to BMP-12. BMC Cell Biology 10(1): 1–10. DOI: https://doi.org/10.1186/1471-2121-10-29

Weingartl H, Sabara M, Pasick J, Van Moorlehem E and Babiuk L. 2002. Continuous porcine cell lines developed from alveolar macrophages: partial characterization and virus susceptibility. Journal of Virological Methods 104(2): 203–16. DOI: https://doi.org/10.1016/S0166-0934(02)00085-X

Weiss M L and Troyer D L. 2006. Stem cells in the umbilical cord. Stem Cell Reviews 2(2): 155–62. DOI: https://doi.org/10.1007/s12015-006-0022-y