Isolation and culture of putative mesenchymal stem cells from equine umbilical cord Wharton’s jelly
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Authors
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N S RATHORE
Assistant Professor, Department of Veterinary Biochemistry, ICAR-National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
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S K KASHYAP
Professor and Head, Department of Veterinary Microbiology and Biotechnology, RAJUVAS, Bikaner
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ANUPAMA DEORA
Teaching Associate, Department of Veterinary Microbiology and Biotechnology, RAJUVAS, Bikaner
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PANKAJ KUMAR
Assitant Disease Investigation Officer, Department of VPHE, LUVAS, Hisar
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J SINGH
Farm Manager, ICAR-National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
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B N TRIPATHI
Director, ICAR-National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
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T R TALLURI
Scientist, Equine Production Campus, ICAR-National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 India
Keywords:
ICAR-National Research Centre on Equines, Jorbeer, Bikaner, Rajasthan 334 001 IndiaAbstract
Despite major progress and knowledge related to the application of adult stem cells, finding alternative sources for bone marrow MSCs has remained a challenge in both humans and animals. In the current study, two protocols namely sequential enzymatic tissue digestion and tissue explant techniques were tried for successful establishment of MSC culture. Umbilical tissues were isolated each time of foaling from five sequential foalings of Marwari mares. Total cell yield, their growth potential and cryopreservation potential were studied. Adherent cell colonies could be established using both isolation methods. Both the cell populations yielded from different protocols performed similarly in terms of population doubling and CFU number value. Additionally, the cells proliferated vigourously and displayed a similar morphology of mesenchymal stem cells. The MSCs were plastic adherent, colonogenic and their morphology was polygonal and fibroblast like. During the proliferation, the cells exhibited density dependent inhibition; analysis of microbial contamination from bacteria, mycoplasma and fungi were negative; the population doubling time of the MSCs isolated was 34.8 h and 40.2 h in enzymatic treatment and tissue explant methods respectively, and diploid chromosome number of the cells was 64, and the diploid frequency was higher than 80%. In conclusion, this study reveals that both the techniques proved to be non-invasive, efficient, simple and quick for isolation and establishment of MSC culture of extra embryonic tissues from equines.
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Arnhold S J, Goletz I, Klein H, Stumpf G, Beluche L A and Rohde C. 2007. Isolation and characterization of bone marrowderived equine mesenchymal stem cells. American Journal of Veterinary Research 68: 1095–105. DOI: https://doi.org/10.2460/ajvr.68.10.1095
Baptista L S, Do Amaral R J, Carias R B, Aniceto M, Claudio-da- Silva C and Borojevic R. 2009. An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples. Cytotherapy 11: 706–15. DOI: https://doi.org/10.3109/14653240902981144
Burk J, Ribitsch I, Gittel C, Juelke H, Kasper C, Staszyk C and Brehm W. 2013. Growth and differentiation characteristics of equine mesenchymal stromal cells derived from different sources. Veterinary Journal 195: 98–106. DOI: https://doi.org/10.1016/j.tvjl.2012.06.004
Cremonesi F, Violini S, Lange C A, Ramelli P, Ranzenigo G and Mariani P. 2008. Isolation, in vitro culture and characterization of foal umbilical cord stem cells at birth. Veterinary Research and Communications 32(Suppl 1): S139–42. DOI: https://doi.org/10.1007/s11259-008-9116-0
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: 1115–22. DOI: https://doi.org/10.1359/jbmr.1999.14.7.1115
Dragoo J, Samimi B, Zhu M, Hame S, Thomas B and Lieberman J. 2003. Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. Journal of Bone Joint Surgery Branch 85(5): 740–47. DOI: https://doi.org/10.1302/0301-620X.85B5.13587
Hefley T, Cushing J and Brand J S. 1981. Enzymatic isolation of cells from bone: cytotoxic enzymes of bacterial collagenase. American Journal of Physiology 240: C234–38. DOI: https://doi.org/10.1152/ajpcell.1981.240.5.C234
Heidari B, Shirazi A, Akhondi M M, Hassanpour H, Behzadi B, Nederi M M, Sarvari A and Borjian S. 2013. Comparison of proliferative and multilineage differentiation potential of sheep mesenchymal stem cells derived from bone marrow, liver and adipose tissue. Avicenna Journal of Medical Biotechnology 5(2): 104–17.
Hoynowski S M, Fry M M, Gardner B M, Leming M T, Tucker J R, Black L, Sand T and Mitchell K E. 2007. Characterization and differentiation of equine umbilical cord derived matrix cells. Biochemical and Biophysical Research Communications 362: 347–53. DOI: https://doi.org/10.1016/j.bbrc.2007.07.182
Huang G T, Gronthos S and Shi S. 2009. Mesenchymal stem cells derived from dental tissues vs. those from other sources. Their biology and role in regenerative medicine. Journal of Dental Research 88(9): 792–06. DOI: https://doi.org/10.1177/0022034509340867
Hyder A. 2005. Effect of the pancreatic digestion with liberase versus collagenase on the yield, function and viability of neonatal rat pancreatic islets. Cell Biology International 29: 831–34. DOI: https://doi.org/10.1016/j.cellbi.2005.05.004
Kasashima Y, Ueno T, Tomita A, Goodship A E and Smith R K. 2011. Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells. Equine Veterinary Journal 43: 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 Orthopedic Research 26: 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 30: 26. 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: 1613–19. DOI: https://doi.org/10.1634/stemcells.2005-0264
Liu K, Wang K and Yan H. 2009. Incomplete digestion preserves chondrocytes from dedifferentiating in long-termed culture on plastic substrate. Tissue and Cell 41: 1–11. DOI: https://doi.org/10.1016/j.tice.2008.06.002
Lovati A B, Corradetti B, Lange C A, 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: 103–21. DOI: https://doi.org/10.1007/s11259-010-9457-3
Patel M, Smith A J, Sloan A J, Smith G and Cooper P R. 2009. Phenotype and behaviour of dental pulp cells during expansion culture. Archives of Oral Biology 54: 898–908. DOI: https://doi.org/10.1016/j.archoralbio.2009.06.008
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: 329–36. DOI: https://doi.org/10.1002/jcp.21312
Rodbell M.1964. Metabolis of isolated fat cells: Effects of hormones on glucose metabolism and lipolysis. Journal of Biological Chemistry 239: 375–80. DOI: https://doi.org/10.1016/S0021-9258(18)51687-2
Sanchez-Guijo F M, Blanco J F, Cruz G, Muntion S, Gomez M, Carrancio S, Lopez-Villar O, Barbado M V, Sanchez-Abarca L I and Blanco B. 2009. Multiparametric comparison of mesenchymal stromal cells obtained from trabecular bone by using a novel isolation method with those obtained by iliac crest aspiration from the same subjects. Cell and Tissue Research 336: 501–07. DOI: https://doi.org/10.1007/s00441-009-0778-x
Talluri T R, Kumar D, Glage S, Garrels W, Ivics Z, Debowski K and Kues W A. 2015. Derivation and characterization of bovine induced pluripotent stem cells by transposon-mediated reprogramming. Cellular Reprogramming 17(2): 131–40. DOI: https://doi.org/10.1089/cell.2014.0080
Toupadakis C A, Wong A, Genetos D C, Cheung WK, 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: 1237–45. DOI: https://doi.org/10.2460/ajvr.71.10.1237
Tsai WC, Tang F T, Hsu C C, Hsu Y H, Pang J H and Shiue C C. 2004. Ibuprofen inhibition of tendon cell proliferation and upregulation of the cyclin kinase inhibitor p21CIP1. Journal of Orthopaedic Research 22: 586–91. DOI: https://doi.org/10.1016/j.orthres.2003.10.014
Vidal M A, Kilroy G E, Lopez M J, Johnson J R, Moore RM and Gimble J M. 2007. Characterization of equine adipose tissuederived stromal cells: adipogenic and osteogenic capacity and comparison with bone marrow-derived mesenchymal stromal cells. Veterinary Surgery 36: 613–22. DOI: https://doi.org/10.1111/j.1532-950X.2007.00313.x
Vidal M A, Walker N J, Napoli E and Borjesson D L. 2012. Evaluation of senescence in mesenchymal stem cells isolated from equine bone marrow, adipose tissue, and umbilical cord tissue. Stem Cells and Development 21: 273–83. DOI: https://doi.org/10.1089/scd.2010.0589
Williams S K, McKenney S and Jarrell B E. 1995. Collagenase lot selection and purification for adipose tissue digestion. Cell Transplantation 4: 281–89. DOI: https://doi.org/10.1177/096368979500400306
Weingartl H M, 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: 203–16. DOI: https://doi.org/10.1016/S0166-0934(02)00085-X
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