Development of a cryopreservation protocol for equine mesenchymal stem cells obtained from umbilical cord

• DOI: 10.1016/j.jevs.2013.10.167
• Publication Date: 2013-12-17T00:46:06Z
• Authors: L. Maia; C.N. Moraes; C.P. Freitas-Dell...; C.M. Queiroz; L.S.L.S. Mota; V. Santiloni; F.C. Landim-Alvar...
• Keywords: 0303 health sciences; 03 medical and health sciences; 3. Good health

In equine medicine, mesenchymal stem cells (MSCs) from bone marrow and adipose tissue are the most common types of adult stem cells (ASC) used for therapeutic approach (VIDAL, M.A. et al. Vet. Surg.,2008; 37:713-24. However, MSC from other sources such as the umbilical cord (UC) have aroused great interest because they share differentiation characteristics between embryonic stem cells and multipotent ASC (CREMONESI, F. et al. Theriogenology,2011; 75:1400-15.The aim of the present work was to evaluate by flow cytometry (FC) the rates of viability and apoptosis in MSC obtained from the umbilical cord Wharton’s jelly matrix (MSC-UC) after cryopreservation with different mediums. For this, samples of UC (n1⁄42) were collected at delivery, processed and cultured in DMEM high glucose supplemented with 20% fetal bovine serum, 1% penicillin/streptomycin, and 1.2% amphotericin B (Invitrogen ) at 37.5 C in a humidified atmosphere, containing 5% CO2 in air. Prior to cryopreservation MSC-UC were characterized showing positive response to adipogenic and osteogenic differentiation, expression of cell surface markers CD44 and intracellular OCT4, absence of MHCII marker, good response to CFU-F assay and normal karyotype (2n1⁄464). MSC were cryopreserved in a controlled freezing system using Mr frosty (-1 C/minute until -80 C -24 hours, Nalgene ) with initial viability of 90 7.42% according to the following protocols: Medium 1: DMEM high glucose, with 20% fetal bovine serum (FBS) and