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使用即用型培养基测量从人多能干细胞获得的心肌细胞中钙尖峰的方案。

Protocol to measure calcium spikes in cardiomyocytes obtained from human pluripotent stem cells using a ready-to-use media.

作者信息

Astro Veronica, Ramirez-Calderon Gustavo, Adamo Antonio

机构信息

Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.

Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.

出版信息

STAR Protoc. 2023 Apr 13;4(2):102252. doi: 10.1016/j.xpro.2023.102252.

DOI:10.1016/j.xpro.2023.102252
PMID:37060558
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10140149/
Abstract

The derivation of cardiomyocytes from human pluripotent stem cells (hPSCs) is a powerful tool to investigate early cardiogenesis and model diseases in vitro. Here, we present an optimized protocol to obtain contracting hPSCs-derived cardiomyocytes using a ready-to-use kit. We describe steps for hPSC culture and differentiation to cardiomyocytes including the identification of key parameters such as starting cell confluency and temperature. We then detail immunofluorescence, flow cytometry, and the quantification of cardiomyocytes' calcium spikes using live imaging. For complete details on the use and execution of this protocol, please refer to Astro et al..

摘要

从人多能干细胞（hPSC）中衍生心肌细胞是研究早期心脏发生和体外疾病建模的有力工具。在此，我们展示了一种使用即用型试剂盒获得收缩性hPSC衍生心肌细胞的优化方案。我们描述了hPSC培养和分化为心肌细胞的步骤，包括确定关键参数，如起始细胞汇合度和温度。然后，我们详细介绍了免疫荧光、流式细胞术以及使用实时成像对心肌细胞钙瞬变进行定量分析的方法。有关本方案使用和执行的完整详细信息，请参考阿斯特罗等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/862b807e61cf/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/9edfd14e1d3f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/4d173da73afd/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/a17bd9b17475/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/744c1aeee4b8/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/6a884a5637be/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/591c0241ec08/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/40a7f46190f4/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/862b807e61cf/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/9edfd14e1d3f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/4d173da73afd/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/a17bd9b17475/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/744c1aeee4b8/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/6a884a5637be/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/591c0241ec08/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/40a7f46190f4/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfd/10140149/862b807e61cf/gr7.jpg

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