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新生小鼠心肌细胞分离培养方案。

Protocol for isolating and culturing neonatal murine cardiomyocytes.

作者信息

Bongiovanni Chiara, Miano Carmen, Sacchi Francesca, Da Pra Silvia, Del Bono Irene, Boriati Stefano, D'Uva Gabriele

机构信息

Department of Medical and Surgical Sciences, University of Bologna, Via Massarenti 9, 40138 Bologna, Italy; Centre for Applied Biomedical Research (CRBA), University of Bologna, Via Massarenti 9, 40138 Bologna, Italy.

Department of Medical and Surgical Sciences, University of Bologna, Via Massarenti 9, 40138 Bologna, Italy; IRCCS Azienda Ospedaliero-Universitaria di Bologna, Via Massarenti 9, 40138 Bologna, Italy.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103461. doi: 10.1016/j.xpro.2024.103461. Epub 2024 Nov 22.

Abstract

The isolation and culture of neonatal murine cardiac cells are valuable techniques for studying their properties and molecular mechanisms in response to various treatments or conditions. Here, we present a protocol for isolating a high yield of viable neonatal murine cardiac cells, including functional, beating cardiomyocytes. We describe the steps of heart extraction, washing and pre-digestion, digestion, and cell seeding. We detail procedures for mechanical and enzymatic digestions, conducted in a controlled environment within a cell culture incubator. For complete details on the use and execution of this protocol, please refer to Bongiovanni et al..

摘要

新生小鼠心脏细胞的分离和培养是研究其在各种处理或条件下的特性和分子机制的重要技术。在此,我们介绍一种分离高产率存活新生小鼠心脏细胞的方法,包括功能性跳动心肌细胞。我们描述了心脏提取、洗涤和预消化、消化以及细胞接种的步骤。我们详细说明了在细胞培养箱的受控环境中进行的机械和酶消化程序。有关本方法的使用和执行的完整详细信息,请参考邦焦万尼等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1735/11625229/b4753e992b69/fx1.jpg

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