Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Abstract

The objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also cultured in specific media for adipogenic and chondrogenic differentiation. There was no expression of the CD13 marker, but CD44 and CD90 were expressed in all of the passages tested. After 14 days of cell differentiation into adipocytes, lipid droplets were observed upon Oil Red O (ORO) staining. Twenty-one days after chondrogenic differentiation, the cells were stained with Alcian Blue. Although the technique for the isolation of these cells requires improvement, the present study demonstrates the partial characterization of PbMSCs, classifying them as a promising type of progenitor cells for use in equine cell therapy.

Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells

“…Prior to cryopreservation, UCIM-MSCs were satisfactorily characterized with respect to morphology, plastic adherence, immunophenotypic profile, potential to differentiate into mesodermal lineages, clonicity, and chromosomal stability. These characteristics corroborate with some findings observed in other studies with UCIM-MSCs from umbilical cord (Hoynowski et al, 2007;Lange-Consiglio et al, 2011;Barberini et al, 2014), amniotic liquid (De , amniotic membrane (Lange-Consiglio et al, 2012), bone marrow (Maia et al, 2012(Maia et al, , 2015, adipose tissue (Carvalho et al, 2013b), and peripheral blood (Martinello et al, 2010;Carvalho et al, 2013a).…”

Tratamiento de la laminitis crónica en equinos utilizando células troncales mesenquimales alogénicas de la médula ósea

“…ABM-MSC used for infusion were positive to the surface markers CD73, CD90, CD105, CD28 and CD44 (CD28/CD44 specific for equine MSC), negative to CD45, CD34, CD14 and CD79. Flow cytometry was conducted based upon protocols previously reported (24)(25)(26)(27)(28) .…”