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Cellular Characteristics of Cultured Canine Trophoblasts
Author(s) -
Sahlfeld L,
Hazzard T,
Kutzler M
Publication year - 2012
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12060
Subject(s) - trophoblast , biology , cytotrophoblast , decidua , matrix metalloproteinase , placentation , cytokeratin , immunocytochemistry , andrology , placenta , collagenase , cell culture , microbiology and biotechnology , pathology , immunohistochemistry , immunology , fetus , endocrinology , pregnancy , enzyme , medicine , biochemistry , genetics
Contents Shallow trophoblast invasion is detrimental in human pregnancies, but represents normal endotheliochorial placentation in dogs. Factors regulating shallow trophoblast invasion into the canine decidua are not well described, but it is known that matrix metalloproteinases ( MMP s) play a crucial role in trophoblast invasion in many species. Following the methods previously described for isolating human trophoblasts, canine trophoblasts were isolated using collagenase and trypsin digestions with P ercoll density gradient centrifugation. In addition, placental pieces were cryopreserved prior to primary culture following methods previously described for human tissue. Expression of cytokeratin‐7, MMP 2 and MMP 9 was confirmed using fluorescent immunocytochemistry. Cellular morphology was similar to that reported for trophoblasts. More than 97% of the cells cultured expressed cytokeratin‐7. More cultured canine trophoblasts expressed MMP 9 (54.7 ± 3.4%) compared with MMP 2 (40.3 ± 1.8%) (p = 0.02). Although both MMP s were immunolocalized to the cytoplasm, MMP 2 was found in large, coalescing granules, whereas MMP 9 was more diffusely expressed throughout the cell. Cryogenic freezing of placental tissue prior to primary cell culture had no effect on cell proliferation (p = 0.37). This research has established a baseline for future studies investigating the canine placenta as a model for disorders of shallow trophoblast invasion in humans.