Isolation and characterization of stem cells derived from ovarian tissue of adult dogs

• Grant number: 13/14293-0
• Support Opportunities: Scholarships in Brazil - Master
• Effective date (Start): December 01, 2013
• Effective date (End): December 31, 2014
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Joaquim Mansano Garcia
• Grantee: Amanda Baracho Trindade Hill
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The study of canine stem cells contributes with the progress of veterinary and human cellular therapies, because the dog is often the best animal model for many hereditary and acquired human disease. Due to their capacity for self-renewal and differentiation into multiple cell types, stem cells (SC) are considered the next frontier for regenerative medicine. While embryonic SC have unlimited differentiation potential (i.e.pluripotent), adult-derived multipotent SC are often preferred for ease of recovery from the patient itself (i.e. autologous) and for avoiding ethical constraints of using embryos. Although bone marrow- derived SC were the first and most extensively studied in adult, MSC isolated from adipose tissue have shown several advantages, including easier cell isolation and greater stromal cell density. Nonetheless, identifying new sources of adult SC is essential for scientific progress in cellular therapy. For years it was postulated that female germ cells do not have the potential of self-renewal after birth, but recent promising studies in mice and humans have shown the existence SC in ovarian tissue, that have greater proliferative and differentiation potential than SC in other adult tissues. Based on this evidence, the aim of this project is to obtain stem cells isolated from ovarian tissue of adult dogs. In experiment I, we will evaluate the collagenase- digested for ovarian- and adipose derived SC. For each tissue type we will evaluate the number of viable cells, colony morphology, time to reach confluence, population-doubling, cryotolerance, senescence and cytogenic stability. Experiment II is be aimed at further characterizing at the molecular level the SC obtained in Exp.I. We will evaluate alkaline phosphatase activity, expression of mesenchymal markers through immunofluorescence (flow cytometry and confocal microscopy) and RT-PCR and the ability to differ into adipogenic, osteogenic and condrogenic tissue. Overall, we expect to identify an ovarian source of adult SC for a wide range of clinical applications. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

TRINDADE, A. B.; THERRIEN, J.; GARCIA, J. M.; SMITH, L. C.. Mesenchymal-like stem cells in canine ovary show high differentiation potential. Cell Proliferation, v. 50, n. 6, DEC 2017. (13/14293-0)

HILL, AMANDA B. T.; HILL, JONATHAN E. B. T.; BRESSAN, FABIANA F.; MIGLINO, MARIA ANGELICA; GARCIA, JOAQUIM M.. Derivation and Differentiation of Canine Ovarian Mesenchymal Stem Cells. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, n. 142, DEC 2018. (13/14293-0)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

HILL, Amanda Baracho Trindade. Derivação de células tronco de tecido ovariano na espécie canina. 2014. Master's Dissertation - Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal Jaboticabal.