Successful isolation and culture of equine placental cells from failed early pregnancies

Abstract

Early pregnancy loss (EPL) prior to day 65 occurs in 610% of all equine pregnancies and accounts for over 50% of Thoroughbred pregnancies lost in the UK. Despite the clear influence of this condition on fertility, we still know relatively little about the causes of EPL. We hypothesise that a proportion of cases of EPL, in particular in older mares, arise due to chromosomal defects such as trisomies and more rarely translocations. Progress in identifying genetic cause(s) of EPL has been hampered due to a reported lack of failed conceptus tissue and a method to isolate and culture cells obtained from those conceptuses. The objective of this study was to develop a method to isolate and culture placental cells isolated from failed pregnancies by comparing the viability and culture success rates of placental cells cultured in different media and processed at different time intervals following uterine lavage. All Thoroughbred mares included in the study were housed on studfarms in the Newmarket area, UK. Conceptus material from pregnancies that failed prior to day 65 was isolated by sterile uterine lavage performed by the attending veterinary surgeon. Conceptuses were transported to the laboratory on ice in HBSS, 5% Equine Serum, 2.5 mg/ml amphotericin, 20 ng/ml kanamycin and 100 U/ml penicillin-streptomycin. Tissue was isolated from the allantochorion, chorion and chorionic girdle (if present) and cells cultured in three different media; AmnioChrome II,