Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium

DOI: 10.18805/IJAR.B-4494    | Article Id: B-4494 | Page : 429-433
Citation :- Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium.Indian Journal of Animal Research.2022.(56): 429-433
Ajeet Kumar Jha, Anirban Mandal, Kalyani Ray, Shyamal Kanti Guha ajitjha2003@gmail.com
Address : Department of Veterinary Gynaecology and Obstetrics, West Bengal University of Animal and Fishery Sciences, Kolkata-700 037, West Bengal, India.
Submitted Date : 23-04-2021
Accepted Date : 9-06-2021


Background: Few studies have confirmed the presence of ovarian tissue stem cells indicating the capacity for differentiation. Based on this fact, it was hypothesized that mesenchymal stem cells (MSC) were found in ovarian surface epithelium (OSE) of canines that could easily be isolated. 
Methods: Both left and right ovaries were minced and digested using collagenase to obtain a stromal vascular fraction (SVF). MSCs were characterized using RT-PCR. To ascertain the trilineage differentiation potential, MSCs were stained with respective stain for osteocytes, chondrocytes and adipocytes. 
Result: We observed elongated, spindle-shaped and fibroblast like appearance of cells after 72 h of initial culture. Expression of MSC specific surface markers were observed through RT-PCR. Using Stem Pro® differentiation medium, OSE were differentiated into osteogenic, chondrogenic and adipogenic lineages and were found to be potential source for isolation, characterization and differentiation of MSCs. Canine (OSE) is easily accessible, multipotent and has high plasticity, holding promise for applications in regenerative medicine.


Canine ovarian surface epithelium Mesenchymal stem cells Ovariectomy Stromal vascular fraction Trilineage differentiation


  1. Auersperg, N., Wong, A.S., Choi, K., Kang, S.K. and Leung, P.C.K. (2001). Ovarian Surface Epithelium/: Biology, Endocrinology. Endocriology. 22(2): 255-288.
  2. Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Keating, A., Prockop, D. and Horwitz, E. (2006). Minimal criteria for defining multipotent mesen-chymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 8(4): 315-317.
  3. Erices, A., Conget, P. and Minguell, J.J. (2000). Mesenchymal progenitor cells in human umbilical cord blood. British Journal of Haematology. 109(1): 235-42.
  4. Haasters, F., Prall, W.C., Anz, D., Bourquin, C., Pautke, C., Endres, S., Mutschler, W., Docheva, D. and Schieker, M. (2009). Morphological and immunocytochemical characteristics indicate the yield of early progenitors and represent a quality control for human mesenchymal stem cell culturing. Journal of Anatomy. 214(5): 759-767. 
  5. Hill, A.B.T., Hill, J.E.B.T., Bressan, F.F., Miglino, M.A. and Garcia, J.M. (2018). Derivation and Differentiation of Canine Ovarian Mesenchymal Stem Cells. Journal of Visualized Experiments. 142: 1-6. 
  6. Horn, P.A., Morris, J.C., Neff, T. and Kiem, H.P. (2004). Stem cell gene transfer - Efficacy and safety in large animal studies. Molecular Therapy. 10(3): 417-431. 
  7. Johnson, J., Canning, J., Kanedo, T., Pru, J.K. and Tilly, J.L. (2004). Germline Stem Cells and Follicular Renewal in the Postnatal Mammalian Ovary. Obstetrical and Gyneco-logical Survey. 59(7): 518-520.
  8. Kern, S., Eichler, H., Stoeve, J., Klüter, H. and Bieback, K. (2006). Comparative Analysis of Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Blood, or Adipose Tissue. Stem Cells. 24(5): 1294-1301 
  9. Kerr, C.L., Hill, C.M., Blumenthal, P.D. and Gearhart, J.D. (2008). Expression of pluripotent stem cell markers in the human fetal ovary. Human Reproduction. 23: 589-599.
  10. Mandal, A., Jha, A.K., Biswas, D. and Guha, S.K. (2021). Derivation, characterization and in vitro cell regeneration of canine white adipose tissue-derived mesenchymal stem cells obtained from a mesenteric region. The Journal of Basic and Applied Zoology. 82(29): https://doi.org/10.1186/s41936-021-00222-1.
  11. Neupane, M., Kiupel, M. and Yuzbasiyan-gurkan, V. (2008). Isolation and Characterization of Canine Adipose–Derived Mesenchymal Stem Cells. Tissue Engineering Part A. 14(6): 1007-1015.
  12. Ogawa, R., Mizuno, H., Watanabe, A., Migita, M., Shimada, T. and Hyakusoku, H. (2004). Osteogenic and chondrogenic differentiation by adipose-derived stem cells harvested from GFP transgenic mice. Biochemical and Biophysical Research Communications. 313(4): 871-877.
  13. Romanov, Y.A. (2003). Searching for Alternative Sources of Postnatal Human Mesenchymal Stem Cells: Candidate MSC-Like Cells from Umbilical Cord. Stem Cells. 21(1): 105-110.
  14. Salamanca, C.M., Maines-Bandiera, S.L., Leung, P.C. K., Hu, Y.L. and Auersperg, N. (2004). Effects of epidermal growth factor/hydrocortisone on the growth and differentiation of human ovarian surface epithelium. Journal of the Society for Gynecologic Investigation. 11(4): 241-251. 
  15. Stimpfel, M., Cerkovnik, P. and Novakovic, S. (2014). Putative mesenchymal stem cells isolated from adult human ovaries. Journal of Assisted Reproduction and Genetics. 31: 959-974.
  16. Uezumi, A., Ojima, K., Fukada, S. I., Ikemoto, M., Masuda, S., Miyagoe-Suzuki, Y. and Takeda, S. (2006). Functional heterogeneity of side population cells in skeletal muscle. Biochemical and Biophysical Research Communications. 341(3): 864-873.
  17. Wagner, W., Roderburg, C., Wein, F., Diehlmann, A., Frankhauser, M., Schubert, R., Eckstein, V. and Ho, A.D. (2007). Molecular and Secretory Profiles of Human Mesenchymal Stromal Cells and Their Abilities to Maintain Primitive Hematopoietic Progenitors. Stem Cells. 25 (10): 2638-2647.
  18. Wang, H.-S., Hung, S.-C., Peng, S.-T., Huang, C.-C., Wei, H.-M., Guo, Y.-J., Fu, Y.-S., Lai, M.-C. and Chen, C.C. (2004). Mesenchymal Stem Cells in the Wharton’s Jelly of the Human Umbilical Cord. Stem Cells. 22(7):1330-1337.
  19. Yaneselli, K.M., Rial, A., Magrisso, A.B., Kuhl, C.P., de Oliveira, F.S., Cirne-Lima, E., Torman, V., Terraciano, P. B., Llambí, S., Pizzato, S. B., Moreno, M., Pazza, K. and Maisonnave, J. (2018). Comparison of the characteristics of canine adipose tissue-derived mesenchymal stem cells extracted from different sites and at different passage numbers. Journal of Veterinary Science. 19(1): 13-20.
  20. Zajdel, A., Ka³ucka, M., Kokoszka-miko³aj, E. and Wilczok, A. (2017). Osteogenic differentiation of human mesenchymal stem cells from adipose tissue and Wharton’s jelly of the umbilical cord. Acta Biochimica Polonica. 64(2): 365-369.

Global Footprints