title: Differential apoptotic staining of mammalian blastocysts based on double immunofluorescent CDX2 and active caspase-3 staining.
abstract: Several approaches have been described for differential staining of blastocysts, but these methods are often time-consuming and unreliable. Here we describe a method for simultaneous differential staining and detection of apoptosis. The differential staining is based on the transcription factor CDX2 which is localized in the nucleus of trophectoderm (TE) cells but absent in the inner cell mass (ICM). Apoptosis is detected by staining of active caspase-3, a key player in several apoptotic pathways. This new approach represents a robust method for quantifying simultaneously ICM/TE ratio and apoptotic cell ratio in bovine, murine, porcine, and human blastocysts.
authors.affiliation: Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium. Eline.Wydooghe@ugent.be
authors.forename: Eline
authors.initials: E
authors.lastname: Wydooghe
authors.forename: Leen
authors.initials: L
authors.lastname: Vandaele
authors.forename: Josine
authors.initials: J
authors.lastname: Beek
authors.forename: Herman
authors.initials: H
authors.lastname: Favoreel
authors.forename: Björn
authors.initials: B
authors.lastname: Heindryckx
authors.forename: Petra
authors.initials: P
authors.lastname: De Sutter
authors.forename: Ann
authors.initials: A
authors.lastname: Van Soom
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D017209:Apoptosis; D001755:Blastocyst / Q000378:metabolism*; D000071616:CDX2 Transcription Factor; D053148:Caspase 3 / Q000032:analysis* / Q000276:immunology; D002417:Cattle; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000737:chemistry*; D018398:Homeodomain Proteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D051379:Mice; D013194:Staining and Labeling / Q000379:methods*; D013552:Swine
chemical_list: D000906:Antibodies; D000071616:CDX2 Transcription Factor; C491270:CDX2 protein, human; D005456:Fluorescent Dyes; D018398:Homeodomain Proteins; D053148:Caspase 3