人类多能干细胞衍生的肾类器官感染 SARS-CoV-2 的方案。
Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells.
机构信息
Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain.
Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany; Research Group "Cellular Polarity and Viral Infection", German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101872. doi: 10.1016/j.xpro.2022.101872. Epub 2022 Nov 7.
This protocol presents the use of SARS-CoV-2 isolates to infect human kidney organoids, enabling exploration of the impact of SARS-CoV-2 infection in a human multicellular in vitro system. We detail steps to generate kidney organoids from human pluripotent stem cells (hPSCs) and emulate a diabetic milieu via organoids exposure to diabetogenic-like cell culture conditions. We further describe preparation and titration steps of SARS-CoV-2 virus stocks, their subsequent use to infect the kidney organoids, and assessment of the infection via immunofluorescence. For complete details on the use and execution of this protocol, please refer to Garreta et al. (2022)..
本方案介绍了使用 SARS-CoV-2 分离株感染人类肾类器官，从而在人类多细胞体外系统中探索 SARS-CoV-2 感染的影响。我们详细说明了如何从人类多能干细胞 (hPSC) 生成肾类器官，并通过使类器官暴露于类似糖尿病的细胞培养条件来模拟糖尿病环境。我们进一步描述了 SARS-CoV-2 病毒储备的制备和滴定步骤，随后将其用于感染肾类器官，并通过免疫荧光法评估感染情况。如需了解本方案使用和实施的完整详细信息，请参考 Garreta 等人 (2022)。
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