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A fluorescent staining technique for studying vascularity and angiogenesis in interdigitated maternal and fetal villi of sheep placenta
Author(s) -
Redmer Dale A,
Dorsam Sheri T,
GrazulBilska Anna T,
Borowicz Pawel P
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.688.6
Subject(s) - placenta , staining , cd31 , trophoblast , fetus , angiogenesis , intervillous space , pathology , lectin , vascularity , biology , antigen , andrology , microbiology and biotechnology , immunohistochemistry , chemistry , immunology , medicine , pregnancy , genetics , cancer research
The objective of this study was to improve detection of the blood vessel network in placenta while simultaneously distinguishing maternal from fetal villi for image analysis. Therefore, we developed a technique for dual fluorescent labeling of endothelial cells and fetal trophoblast. Placentomes (d90 and d130 of pregnancy) were fixed (10% formalin or Carnoy's soln) and paraffin embedded. Following antigen retrieval (50mM glycine, 1mM EDTA, 0.05% Tween 20, pH 9.0; 120 C, 10 min), maternal and fetal blood vessels were identified by immunofluorescent labeling with an antibody (c/n:ab28364, ABCAM) against CD31 antigen, and CF 633 conjugated IgG (c/n:20122, Biotium). The trophoblast layer was stained with FITC labeled BS1 lectin (c/n:FL‐1101; Vector; 20 μg/ml). CD31 labeling resulted in bright fluorescent localization of all blood vessels. Positive lectin staining of fetal trophoblast allowed differentiation of fetal and maternal compartments. Thus, CD31 staining along with fluorescently labeled lectin combined with microscopy and imaging techniques provide an excellent tool to study angiogenesis and vascular architecture in sheep placenta, and other tissues. Grants: Hatch ND1748; USDA‐NIFA 2011–67016‐30174.