title: Isolation and characterization of equine amnion mesenchymal stem cells.
abstract: The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth kinetic of EAMSCs we have taken into account some parameters [PD (population doubling), fold increase and DT (doubling time)]. The differentiation in chondrogenic, adipogenic and osteogenic types of cells and their epitope expression by a cytofluorimetric study have been reported. EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation. By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.
authors.affiliation: Department of Animal Pathology, University of Pisa, Pisa, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Coli
authors.forename: Francesca
authors.initials: F
authors.lastname: Nocchi
authors.forename: Roberta
authors.initials: R
authors.lastname: Lamanna
authors.forename: Mariacarla
authors.initials: M
authors.lastname: Iorio
authors.forename: Simone
authors.initials: S
authors.lastname: Lapi
authors.forename: Patrizia
authors.initials: P
authors.lastname: Urciuoli
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Scatena
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Giannessi
authors.forename: Maria Rita
authors.initials: MR
authors.lastname: Stornelli
authors.forename: Simona
authors.initials: S
authors.lastname: Passeri
keywords: AM, amniotic membrane; CFU, colony-forming unit; CFU-F, CFU-fibroblast; CPD, cumulative population doubling; DT, doubling time; EAMSC, equine amnion mesenchymal stem cell; EGF, epidermal growth factor; FCS, foetal calf serum; H/E, haematoxylin and eosin; HS, horse serum; MSC, mesenchymal stem cell; Oct-4, octamer-binding transcription factor 4; P/S, penicillin/streptomycin; P0, passage 0; PD, population doubling; amnion; c-Kit, cluster tyrosine-protein kinase; equine; growth kinetics; histochemistry; immunohistochemistry; mesenchymal stem cells (MSCs); α-MEM, α-modified minimum essential medium