Introduction: Successful implantation and placentation are essential for the normal intrauterine development of the fetus. Trophoblast cell proliferation, differentiation, invasive behaviour and interaction with the maternal immune system are known to have an impact on these processes. Trophoblast cells do not only physically anchor the developing fetus to the uterus, but they give rise to the syncytiotrophoblast. This is the principal endocrine component of the placenta, and participates in essential metabolic processes and in the defence against transplacentally transmitted infections. Therefore, placental trophoblasts play an important role in the establishment and maintenance of pregnancy.
Aim of the study: The authors summarize their experience with the isolation, characterization and culture of cytotrophoblast cells from first-trimester human placentae.
Materials and methods: The simultaneous preparation of highly enriched human placental trophoblast populations from first-trimester placental villi (6-12 weeks of gestation) by sequential trypsinization, Percoll gradient centrifugation, and negative selection with anti-CD45 or HLA-ABC and HLA-DP, DQ, DR immunomagnetic separation is described. Characterization of freshly isolated and cultured cytotrophoblast cells has been performed by immunohistochemistry, immunofluorescent staining, measurement of hCG production and analysis of matrix metalloproteinase production by substrate gel zymography.
Results and conclusion: On the basis of immunohistochemical and functional data the isolated cells proved to be cytotrophoblasts. This method of isolation and cultivation should facilitate in vitro studies of trophoblast differentiation, invasion, endocrine function, virus interaction, and immunology.