title: A method for isolating and culturing placental cells from failed early equine pregnancies.
abstract: Early pregnancy loss occurs in 6-10% of equine pregnancies making it the main cause of reproductive wastage. Despite this, reasons for the losses are known in only 16% of cases. Lack of viable conceptus material has inhibited investigations of many potential genetic and pathological causes. We present a method for isolating and culturing placental cells from failed early equine pregnancies. Trophoblast cells from 18/30 (60%) failed equine pregnancies of gestational ages 14-65 days were successfully cultured in three different media, with the greatest growth achieved for cells cultured in AmnioChrome™ Plus. Genomic DNA of a suitable quality for molecular assays was also isolated from 29/30 of these cases. This method will enable future investigations determining pathologies causing EPL.
authors.affiliation: Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom.
authors.forename: B V
authors.initials: BV
authors.lastname: Rose
authors.affiliation: Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom.
authors.forename: V
authors.initials: V
authors.lastname: Cabrera-Sharp
authors.affiliation: Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom.
authors.forename: M J
authors.initials: MJ
authors.lastname: Firth
authors.affiliation: Rossdales Equine Practice, Beaufort Cottage Stables, High Street, Newmarket, Suffolk CB8 8JS, United Kingdom.
authors.forename: F E
authors.initials: FE
authors.lastname: Barrelet
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: S
authors.initials: S
authors.lastname: Bate
authors.affiliation: Rossdales Equine Practice, Beaufort Cottage Stables, High Street, Newmarket, Suffolk CB8 8JS, United Kingdom.
authors.forename: I J
authors.initials: IJ
authors.lastname: Cameron
authors.affiliation: Equine Reproductive Services (UK) Ltd., 33 Westgate, Old Malton, Malton, North Yorkshire, YO17 7HE, United Kingdom.
authors.forename: J R
authors.initials: JR
authors.lastname: Crabtree
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Crowhurst
authors.affiliation: Rossdales Equine Practice, Beaufort Cottage Stables, High Street, Newmarket, Suffolk CB8 8JS, United Kingdom.
authors.forename: A J
authors.initials: AJ
authors.lastname: McGladdery
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: H
authors.initials: H
authors.lastname: Neal
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Pynn
authors.affiliation: Rossdales Equine Practice, Beaufort Cottage Stables, High Street, Newmarket, Suffolk CB8 8JS, United Kingdom.
authors.forename: O D
authors.initials: OD
authors.lastname: Pynn
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: C
authors.initials: C
authors.lastname: Smith
authors.affiliation: Newmarket Equine Hospital, Cambridge Road, Newmarket, Suffolk, CB8 0FG, United Kingdom.
authors.forename: Z
authors.initials: Z
authors.lastname: Wise
authors.affiliation: Department of Production and Population Health, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom.
authors.forename: K L P
authors.initials: KL
authors.lastname: Verheyen
authors.affiliation: Department of Production and Population Health, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom.
authors.forename: D C
authors.initials: DC
authors.lastname: Wathes
authors.affiliation: Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, United Kingdom. Electronic address: ademestre@rvc.ac.uk.
authors.forename: A M
authors.initials: AM
authors.lastname: de Mestre
mesh_terms: D000034:Abortion, Veterinary / Q000000981:diagnostic imaging / Q000473:pathology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods* / Q000662:veterinary; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D020964:Embryo Loss / Q000000981:diagnostic imaging / Q000473:pathology* / Q000662:veterinary; D005260:Female; D005865:Gestational Age; D006736:Horses; D010920:Placenta / Q000000981:diagnostic imaging / Q000473:pathology*; D011247:Pregnancy; D014327:Trophoblasts / Q000473:pathology; D016216:Ultrasonography, Prenatal / Q000662:veterinary
keywords: Cell culture; Embryonic loss; Equine; Pregnancy loss; Trophoblast