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人诱导多能干细胞分化滋养层细胞的胎盘芯片模型建立的研究方案

Protocol for a placenta-on-a-chip model using trophoblasts differentiated from human induced pluripotent stem cells.

机构信息

Systems Medicine, School of Medicine, University of Dundee, DD1 9SY Dundee, UK.

MIMETAS, 2342 DH, Oegstgeest, the Netherlands.

出版信息

STAR Protoc. 2024 Mar 15;5(1):102879. doi: 10.1016/j.xpro.2024.102879. Epub 2024 Feb 14.

Abstract

Organ-on-a-chip technology incorporating stem cell techniques represents a promising strategy to improve modeling of human organs. Here, we present a protocol for generating a standardized 3D placenta-on-a-chip model using trophoblast derived from human induced pluripotent stem cells (hiPSCs). We describe steps for seeding hiPSCs into multi-chip OrganoPlate devices and on-chip differentiation into trophoblasts against an extracellular matrix under perfused conditions. We then detail procedures for conducting a functional barrier integrity assay, immunostaining, and collecting protein or RNA for molecular analysis. For complete details on the use and execution of this protocol, please refer to Lermant et al. (2023)..

摘要

器官芯片技术结合干细胞技术代表了一种改进人类器官建模的有前途的策略。在这里,我们提出了一种使用源自人诱导多能干细胞(hiPSC)的滋养层细胞生成标准化 3D 胎盘芯片模型的方案。我们描述了将 hiPSC 接种到多芯片 OrganoPlate 设备中的步骤,并在灌流条件下在细胞外基质上对其进行向滋养层的分化。然后,我们详细介绍了进行功能屏障完整性测定、免疫染色以及收集蛋白质或 RNA 进行分子分析的步骤。有关该方案的使用和执行的完整详细信息,请参阅 Lermant 等人(2023 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12fe/10876593/7fd857c531ae/fx1.jpg

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