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A Physiological Biomimetic Culture System for Pig and Human Heart Slices

ABSTRACT: A Physiological Biomimetic Culture System for Pig and Human Heart Slices

PROVIDER: PRJNA552335 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9627720_1.fastq.gz	Fastqsanger.gz
	SRR9627720_2.fastq.gz	Fastqsanger.gz
	SRR9627721_1.fastq.gz	Fastqsanger.gz
	SRR9627721_2.fastq.gz	Fastqsanger.gz
	SRR9627722_1.fastq.gz	Fastqsanger.gz

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Project description: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 µm 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 ?-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.

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A Physiological Biomimetic Culture System for Pig and Human Heart Slices

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