{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T13:10:20Z","timestamp":1791378620309,"version":"4.3.2"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,8,6]],"date-time":"2024-08-06T00:00:00Z","timestamp":1722902400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003033","name":"Ministerio de Ciencia, Tecnolog\u00eda e Innovaci\u00f3n","doi-asserted-by":"publisher","award":["PICT 2017-0234, PICT 2020-0862"],"award-info":[{"award-number":["PICT 2017-0234, PICT 2020-0862"]}],"id":[{"id":"10.13039\/501100003033","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Human trophoblastic cell lines, such as BeWo, are commonly used in 2D models to study placental <jats:italic>Trypanosoma cruzi<\/jats:italic> infections. However, these models do not accurately represent natural infections. Three-dimensional (3D) microtissue cultures offer a more physiologically relevant in vitro model, mimicking tissue microarchitecture and providing an environment closer to natural infections. These 3D cultures exhibit functions such as cell proliferation, differentiation, morphogenesis, and gene expression that resemble in vivo conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We developed a 3D culture model using the human trophoblastic cell line BeWo and nonadherent agarose molds from the MicroTissues\u00ae 3D Petri Dish\u00ae system. Both small (12\u2013256) and large (12\u201381) models were tested with varying initial cell numbers. We measured the diameter of the 3D cultures and evaluated cell viability using Trypan Blue dye. Trophoblast functionality was assessed by measuring \u03b2-hCG production via ELISA. Cell fusion was evaluated using confocal microscopy, with Phalloidin or ZO-1 marking cell edges and DAPI staining nuclei. <jats:italic>T. cruzi<\/jats:italic> infection was assessed by microscopy and quantitative PCR, targeting the EF1-\u03b1 gene for <jats:italic>T. cruzi<\/jats:italic> and GAPDH for BeWo cells, using three parasite strains: VD (isolated from a congenital Chagas disease infant and classified as Tc VI), and K98 and Pan4 (unrelated to congenital infection and classified as Tc I).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Seeding 1000 BeWo cells per microwell in the large model resulted in comparable cellular viability to 2D cultures, with a theoretical diameter of 408.68 \u00b1 12.65 \u03bcm observed at 5 days. Functionality, assessed through \u03b2-hCG production, exceeded levels in 2D cultures at both 3 and 5 days. <jats:italic>T. cruzi<\/jats:italic> infection was confirmed by qPCR and microscopy, showing parasite presence inside the cells for all three tested strains. The distribution and progression of the infection varied with each strain.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>This innovative 3D model offers a simple yet effective approach for generating viable and functional cultures susceptible to <jats:italic>T. cruzi<\/jats:italic> infection, presenting significant potential for studying the placental microenvironment.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1433424","type":"journal-article","created":{"date-parts":[[2024,8,6]],"date-time":"2024-08-06T05:12:58Z","timestamp":1722921178000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Novel 3D human trophoblast culture to explore T. cruzi infection in the placenta"],"prefix":"10.3389","volume":"14","author":[{"given":"Sofia","family":"Apodaca","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Di Salvatore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arturo","family":"Mu\u00f1oz-Calder\u00f3n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda de los \u00c1ngeles","family":"Curto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia A.","family":"Longhi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro G.","family":"Schijman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,8,6]]},"reference":[{"key":"B1","first-page":"33","article-title":"Chagas disease in Latin America: an epidemiological update based on 2010 estimates","volume":"90","year":"2015","journal-title":"Wkly Epidemiol. 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