IN VITRO MODEL OF POLARIZED TROPHOBLAST

极化滋养细胞的体外模型

• 批准号: 3147279
• 负责人: JOSEPH DANCIS
• 金额: $ 20.29万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3147279
• 关键词: AIDS; antiAIDS agent; antibody receptor; apical membrane; basolateral membrane; congenital infection; drug adverse effect; drug metabolism; electron microscopy; gene expression; hormone biosynthesis; human immunodeficiency virus; human tissue; immunoglobulin G; inulin; membrane permeability; membrane potentials; model; model design /development; mother /embryo /fetus nutrition; nutrition related tag; placenta; placental transfer; tissue /cell culture; transferrin; trophoblast

An in vitro model of trophoblast will be developed which will permit research into many aspects of placental function as it relates to maternal HIV infection. The trophoblast will be maintained in tissue culture in a bicameral chamber providing independent access to the apical (maternal) and basal (fetal) surfaces of the trophoblast. The criteria for a successful preparation will include tests of barrier function (electrical resistance, restricted transfer of inulin and other molecules, microscopic evidence of a continuous monolayer) and tests of polarity (differentiation into apical and basolateral domains on EM, directed extrusion of virions and preferential uptake of transferrin from the apical surface). Three cell types will be used for the model: transformed trophoblast and primary trophoblast isolated from term and from first trimester placenta. The models will be subjected to tests designed to investigate two of its major roles: the transfer of nutrients to the fetus and the synthesis and secretion of hormones for the maintenance of pregnancy. The potential toxicity of anti-HIV drugs will be explored by exposing trophoblast to the drug and then re-testing its function. The transfer and metabolism of the HIV drugs will also be studied taking advantage of the polarity of the model. Concurrently the usefulness of the model for the study of immunological questions concerning AIDS and other intrapartum infections will be explored. The expression of Fc receptors and the transepithelial transfer of IgG will be measured. Introduction of Fc receptors into trophoblast by recombinant techniques will be undertaken to amplify this function if necessary. The model will be applied to questions pertinent to the manifestation of disease in the fetus such as the transfer of specific antibodies, of viral antigens and infectious agents and of complexes of antigens and antibodies.

将开发一种滋养层的体外模型， 研究胎盘功能的许多方面， 孕产妇艾滋病毒感染。 滋养层将保持在组织中 在两院制培养室中培养，提供独立的根尖通道 （母体）和基底（胎儿）表面的滋养层。 的标准 成功的准备将包括屏障功能的测试 （电阻，菊粉和其他限制转移 分子，连续单层的微观证据）和 极性（在EM上分化为顶端和基底外侧区域， 病毒体的定向挤出和转铁蛋白的优先摄取 顶面）。 模型将使用三种细胞类型： 转化的滋养层和初级滋养层分离自足月妊娠， 从孕早期胎盘中提取的 这些模型将接受测试 旨在研究它的两个主要作用：营养物质的转移 对胎儿和合成和分泌激素的 维持妊娠。 抗艾滋病毒药物的潜在毒性将 通过将滋养层暴露于药物，然后重新测试其 功能 艾滋病毒药物的转移和代谢也将是 利用模型的极性进行研究。 兼任 免疫学问题研究模型的实用性 关于艾滋病和其他产时感染的问题将进行探讨。 的 Fc受体的表达和IgG的跨上皮转移将 被衡量。 Fc受体在滋养层细胞中的表达 将采用重组技术来扩增该功能，如果 必要 该模型将被应用于有关的问题， 胎儿疾病的表现，如特定的 抗体、病毒抗原和感染因子的抗体以及 抗原和抗体。