title: Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse.
abstract: Mesenchymal stem cells (MSCs) have been derived from multiple sources of the horse including umbilical cord blood (UCB) and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), CB and Wharton's Jelly (WJ). Samples were obtained from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was prepared by incubating with an enzymatic solution for 2  h. All cell lines were cultured in DMEM/TCM199 plus fetal bovine serum. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB and 8/12 (66.7%) WJ samples. Statistically significant differences were found between cell-doubling times (DTs): cells isolated from WJ expanded more rapidly (2.0±0.6 days) than those isolated from CB (2.6±1.3 days) and AF (2.3±1.0 days) (P<0.05). Positive von Kossa and Alizarin Red S staining confirmed osteogenesis. Alcian Blue staining of matrix glycosaminoglycans illustrated chondrogenesis and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105; and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from a uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their use as an allogenic cell source, further studies are needed for evaluating the expression of markers related to cell immunogenicity.
authors.affiliation: Department of Veterinary Medical Sciences, University of Bologna, via Tolara di Sopra 50, 40064 Ozzano Emilia, Bologna, Italy. eleonora.iacono2@unibo.it
authors.forename: Eleonora
authors.initials: E
authors.lastname: Iacono
authors.forename: Lara
authors.initials: L
authors.lastname: Brunori
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pirrone
authors.forename: Pasquale Paolo
authors.initials: PP
authors.lastname: Pagliaro
authors.forename: Francesca
authors.initials: F
authors.lastname: Ricci
authors.forename: Pier Luigi
authors.initials: PL
authors.lastname: Tazzari
authors.forename: Barbara
authors.initials: B
authors.lastname: Merlo
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006736:Horses; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011247:Pregnancy; D059631:Wharton Jelly / Q000166:cytology*