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COMPARATIVE STUDY OF CRYOPRESERVATIVE EFFECT ON BIOLOGICAL PROPERTIES OF MESENCHYMAL STEM CELLS FROM THE MATRIX OF UMBILICAL CORD AND BONE MARROW OF HORSES

Grant number: 12/23888-4
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): June 01, 2013
Effective date (End): January 10, 2016
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal researcher:Fernanda da Cruz Landim
Grantee:Leandro Maia
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated scholarship(s):15/17619-9 - RESEARCH TRAINING IN PROTEOMICS:Application of proteomics approaches to evaluate the impact of cryopreservation in the level of expression of proteins in equine mesenchymal stem cells., BE.EP.PD

Abstract

Mesenchymal stem cells (MSCs) have pointed as a promising alternative for cellular therapy because of the large capacity of self-regeneration, low immunogenicity, paracrine, anti-inflammatory, immunomodulatory and antiapoptotic effects. Important properties such as low immunogenicity and absence of reactions observed in allogeneic implants with MSCs, are features that corroborate for the performance of studies that assess the determination of mediums and methods of cryopreservation that allow the creation of a cell bank with high viability rate and maintenance of the biological characteristics after thawing for therapeutic use. In this context, the general aim of this study is to evaluate and compare the effects of cryopreservation with different mediums of freezing on the viability and biological properties of MSCs from bone marrow (MSCs - BM) and matrix of umbilical cord (MSCs - UC). For this, samples of BM and UC of horses will be collected, processed and properly characterized prior to cryopreservation by assessing the ability of formation of fibroblastic colonies (CFUF), the expression or absence of cell surface markers (CD44 +, CD90 +, CD34 - and MHC-II-), assays for differentiation in the tri-lineage (osteogenic, adipogenic and chondrogenic), karyotype analysis and evaluation of quantitative and qualitative proteic profile by proteomic analysis. At the end of the second passage, MSCs previously characterized will be submitted to cryopreservation with four different mediums of freezing in slow freezing system. After three months of cryopreservation samples will be thawed and compared with respect to viability using the markers annexin V and propidium iodide by flow cytometry, beyond the analyses by trypan blue under light microscopy. Additionally will be evaluate the maintenance CUCF, multipotentiality, capacity of expression or no of cell surface markers, occurrence or no of aneuploidy by karyotype analysis and evaluation of proteic profile by proteomic analysis. The results obtained in this study will provide important informations for a better understanding of the biology of MSCs from MO and UC before and after cryopreservation thus favoring the creation of a cell bank from the (s) best (s) source (s) (UC and / or BM) observed (s), using protocol (s) of cryopreservation at low cost, which has as purpose the maintenance of cells with good quality for future studies in vitro and immediate therapeutic application in equine medicine and possibly in other species. Additionally, proteomic analysis of equine fresh and post cryopreservation MSCs provides an innovative character to the work for allowing the identification and better understanding of post-transducional knowledge of different types of proteins (structure, adhesion, transport, signaling, regulation, antioxidant and others) that compose the proteome of MSCs from UC and BM.

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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)
MAIA, LEANDRO; DE MORAES, CAROLINA NOGUEIRA; DIAS, MARIANNE CAMARGOS; BAUZA MARTINEZ, JULIA; ODENA CABALLOL, ANTONIA; TESTONI, GIORGIA; DE QUEIROZ, CARLA MARTINS; DIAZ PENA, RAMON; LANDIM-ALVARENGA, FERNANDA C.; DE OLIVEIRA, ELIANDRE. A proteomic study of mesenchymal stem cells from equine umbilical cord. Theriogenology, v. 100, p. 8-15, SEP 15 2017. Web of Science Citations: 2.
MAIA, LEANDRO; DIAS, MARIANNE CAMARGOS; DE MORAES, CAROLINA NOGUEIRA; FREITAS-DELL'AQUA, CAMILA DE PAULA; SILVEIRA DA MOTA, LIGIA S. L.; SANTILONI, VALQUIRIA; LANDIM-ALVARENGA, FERNANDA DA CRUZ. Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells. Cell Biology International, v. 41, n. 3, p. 239-248, MAR 2017. Web of Science Citations: 1.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.