The term basal plate of the human placenta as a source of functional extravillous trophoblast cells

Background\ud Extravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and several collaborative cross talks involved in establishing and maintaining a successful pregnancy. We report here an improved protocol for the isolation of fully differentiated EVT cells from the basal plate of the human term placenta.\ud \ud \ud Methods\ud The basal plate was carefully dissected from the villous tissue and the amniochorion membrane prior to enzymatic digestion. Term basal EVT cells were isolated using a 30 and 60% Percoll gradient. A panel of markers and characteristics of the isolated cells were used to confirm the specificity and efficiency of the method so that their potential as an investigative tool for placental research could be ascertained.\ud \ud \ud Results\ud Isolated cells were immunoreactive for cytokeratin-7 (CK-7), placental growth factor, placental alkaline phosphatase, human leukocyte antigen G1 (HLA-G1), and α1 and α5 integrins, similarly to the EVT markers from first trimester placental villi. Around 95% of the isolated cells labeled positively for CK-7 and 82% for HLA-G1. No significant change in viability was observed during 48 h of EVT culture as indicated by propidium iodide incorporation and trypan blue test exclusion. Genes for metalloproteinases MMP-2 and MMP9 (positive regulators of trophoblast invasiveness) were expressed up to 48 h of culturing, as also the gelatinolytic activity of the isolated cells. Transforming growth factor (TGF)-beta, which inhibits proliferation, migration, and invasiveness of first-trimester EVT cells, also reduced invasion of isolated term EVT cells in transwell assays, whereas epidermal growth factor was a positive modulator.\ud \ud \ud Conclusions\ud Term basal plate may be a viable source of functional EVT cells that is an alternative to villous explant-derived EVT cells and cell lines. Isolated term EVT cells may be particularly useful in investigation of the role of trophoblast cells in pathological gestations, in which the precise regulation and interactive ability of extravillous trophoblast has been impaired.Fundação de Amparo à Pesquisa do Estado de São Paulo [2009/05354-0; 2013/12243-5]Conselho Nacional de Desenvolvimento Científico e Tecnológico [40088/2010-5]Coordenação de Aperfeiçoamento de Pessoal de Nível Superior [4178-11-4]Austrian Science Funds [P-22687-B13

Changes in the TNF-alpha/IL-10 ratio in hyperglycemia-associated pregnancies

Aims: TNF-alpha is a diabetogenic cytokine associated with adverse outcomes during pregnancy that can be counterbalanced by IL-10. We have investigated IL-10 and TNF-alpha balance at maternal and placental levels in hyperglycemia-associated pregnancies.Methods: One hundred and ninety-two pregnant women participated, which included normoglycemic women (ND) and women with mild gestational hyperglycemia (MGH), gestational diabetes mellitus (GDM) and type 2 diabetes mellitus (DM2). Maternal plasma and placental tissue IL-10 and TNF-alpha levels were measured by ELISA and placental TNF-alpha was also immunolocalized.Results: Maternal plasma TNF-alpha levels were highest in GDM (p = 0.0190), whereas TNF-alpha levels were highest in placental tissues in DM2 (p = 0.0095). Immunohistochemistry also showed strong reactivity with anti-TNF-alpha antibody in the villous structures in the DM2 group. Conversely, IL-10 levels were lowest in maternal plasma of the DM2 group (p = 0.0228). The TNF-alpha/IL-10 ratio in maternal plasma progressively increased with the severity of hyperglycemia (p < 0.0001), being highest in placenta of the DM2 group (p = 0.0150). In both, plasma and placenta, TNF-alpha/IL-10 ratio were correlated with mean maternal glycemia and HbA1c levels.Conclusions: Alterations of placenta and serum TNF-alpha/IL-10 balance with predominance of TNF-alpha were correlated with the severity of hyperglycemia during gestation. This association may offer insight into the pathogenesis of gestational hyperglycemia and associated pregnancy outcomes. (C) 2015 Elsevier Ireland Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

The limited knowledge of placental damage due to neglected infections: ongoing problems in Latin America

The placenta works as a selective barrier, protecting the fetus from potential infections that may affect the maternal organism during pregnancy. In this review, we will discuss several challenging infections that are common within Latin American countries and that may affect the maternal-fetal interface and pose risks to fetal development. Specifically, we will focus on emerging infectious diseases including the arboviruses, malaria, leishmaniasis, and the bacterial foodborne disease caused by Shiga toxinproducing Escherichia coli. We will also highlight some topics of interest currently being studied by research groups that comprise an international effort aimed at filling the knowledge gaps in this field. These topics address the relationship between exposure to microorganisms and placental abnormalities, congenital anomalies, and complications of pregnancy.Fil: Moreti Ribeiro, Isabel. Universidade Federal do Mato Grosso do Sul; BrasilFil: Souza Souto, Paula Cristina. Universidade Federal do Mato Grosso do Sul; BrasilFil: Borbely, Alexandre U.. Universidade Federal de Alagoas; BrasilFil: Lopez Lira Tanabe, Eloiza. Universidade Federal de Alagoas; BrasilFil: Cadavid, Angela. Universidad de Antioquia; ColombiaFil: Alvarez, Angela M.. Universidad de Antioquia; ColombiaFil: Bueno, Julio. Universidad de Antioquia; ColombiaFil: Agudelo, Olga. Universidad de Antioquia; ColombiaFil: Garcia Robles, Reggie. Pontificia Universidad Javeriana; ColombiaFil: Ayala Ramirez, Paola. Pontificia Universidad Javeriana; ColombiaFil: Sacerdoti, Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Szasz, Theodora. Augusta University; Estados UnidosFil: Damiano, Alicia Ermelinda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Escudero, Carlos. Universidad del Bio Bio; ChileFil: Lima, Víctor V.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Giachini, Fernanda R.. Universidade Federal do Mato Grosso do Sul; Brasi