期刊
CIRCULATION RESEARCH
卷 125, 期 6, 页码 628-642出版社
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1161/CIRCRESAHA.119.314996
关键词
calcium; cardiotoxicity; genetic therapy; heart failure; therapeutics
资金
- National Institutes of Health (NIH) [P30GM127607]
- American Heart Association [16POST30960017, 16SDG29950012, 16SFRN31400013]
- NIH [P30GM127607, HL147558, HL116848, HL127240, DK119594, P20GM103436, R01HL131647, R01HL130174, R01ES028268, P01HL78825, UM1HL113530]
- American Diabetes Association Pathway to Stop Diabetes Grant [ADA 1-16-JDF-041]
- British Heart Foundation MBPhD studentship [FS/15/35/31529]
- British Heart Foundation (BHF) Centre for Regenerative Medicine [RM/17/1/33377]
Rationale: Preclinical testing of cardiotoxicity and efficacy of novel heart failure therapies faces a major limitation: the lack of an in situ culture system that emulates the complexity of human heart tissue and maintains viability and functionality for a prolonged time. Objective: To develop a reliable, easily reproducible, medium-throughput method to culture pig and human heart slices under physiological conditions for a prolonged period of time. Methods and Results: Here, we describe a novel, medium-throughput biomimetic culture system that maintains viability and functionality of human and pig heart slices (300 mu m thickness) for 6 days in culture. We optimized the medium and culture conditions with continuous electrical stimulation at 1.2 Hz and oxygenation of the medium. Functional viability of these slices over 6 days was confirmed by assessing their calcium homeostasis, twitch force generation, and response to beta-adrenergic stimulation. Temporal transcriptome analysis using RNAseq at day 2, 6, and 10 in culture confirmed overall maintenance of normal gene expression for up to 6 days, while over 500 transcripts were differentially regulated after 10 days. Electron microscopy demonstrated intact mitochondria and Z-disc ultra-structures after 6 days in culture under our optimized conditions. This biomimetic culture system was successful in keeping human heart slices completely viable and functionally and structurally intact for 6 days in culture. We also used this system to demonstrate the effects of a novel gene therapy approach in human heart slices. Furthermore, this culture system enabled the assessment of contraction and relaxation kinetics on isolated single myofibrils from heart slices after culture. Conclusions: We have developed and optimized a reliable medium-throughput culture system for pig and human heart slices as a platform for testing the efficacy of novel heart failure therapeutics and reliable testing of cardiotoxicity in a 3-dimensional heart model. Visual Overview: An online visual overview is available for this article.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据