title: Bioassay for follicle stimulating activity of equine gonadotropic hormone in mare serum using frozen/thawed transiently transfected reporter cells.
abstract: The objective was to establish a cell line-based bioassay for FSH in horse serum for screening samples with high eCG bioactivity. A cell line (HEK293) was transiently cotransfected with an FSH reporter expression plasmid and a cAMP-responsive β-galactosidase reporter plasmid. Cells were bulk frozen, and thawed for assay purposes. This assay was specific for FSH, with no cross-reaction with LH or insulin-like growth factor-1. Standard curves (eCG) and serum samples from pregnant mares passed parallel line bioassay validity tests (linearity and parallelism). Estimates of bioactivity with this bioassay were highly correlated with estimates obtained with the Steelman-Pohley hCG augmentation assay. The colorimetric end point permitted the use of this assay as a rapid screen for FSH bioactivity without the need for animal use or complex cell culture facilities.
authors.affiliation: Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada.
authors.forename: F
authors.initials: F
authors.lastname: Sahmi
authors.forename: E
authors.initials: E
authors.lastname: Nicola
authors.forename: C A
authors.initials: CA
authors.lastname: Price
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods* / Q000662:veterinary; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D005640:Follicle Stimulating Hormone / Q000032:analysis* / Q000097:blood* / Q000378:metabolism / Q000494:pharmacology; D005615:Freezing; D017930:Genes, Reporter / Q000502:physiology; D006064:Gonadotropins, Equine / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000494:pharmacology; D057809:HEK293 Cells; D006736:Horses / Q000097:blood* / Q000378:metabolism; D006801:Humans; D014162:Transfection
chemical_list: D006064:Gonadotropins, Equine; D005640:Follicle Stimulating Hormone