Are Primary Cultures of Trout (Oncorhynchus mykiss) Ventricular Cardiomyocytes Metabolically Viable?

Abstract

The objectives of this study were to establish a protocol for the isolation of metabolically viable ventricular cardiomyocytes from the rainbow trout and to determine which measures may best reflect viability 24 h after isolation. Cardiomyocytes were isolated by enzymatic digestion and maintained in cell suspension. Viability was assessed using Trypan blue dye exclusion, ATP content, oxygen consumption and lactate dehydrogenase (LDH) leakage into the medium. Viability, assessed by these measures did not significantly change over the time period of this study. ATP content did correlate significantly with oxygen consumption but not with Trypan blue exclusion. We conclude that primary cultured cardiomyocytes remain metabolically viable for at least 24 h after isolation. Also, it appears that ATP content and oxygen consumption most adequately reflect metabolic cell viability. To be confident with a culture, however, a combination of viability measures is necessary when isolating cardiomyocytes from fish.

In vitro expansion of autonomously contracting, cardiomyogenic structures from rainbow trout Oncorhynchus mykiss

“…Already Nurmi & Vornanen (2002) isolated ventricular myocytes from an adult O. mykiss and held them in culture for at least 10 days in serum‐free medium in order to characterize their electrophysiological potential. Goldstein et al (2004) established a protocol for the isolation of metabolically active ventricular cardiomyocytes from O. mykiss , which are viable for at least 24 h after isolation. Furthermore, Brette et al (2008) used a protocol for the isolation of ventricular myocytes from zebrafish Danio rerio (Hamilton) to obtain calcium‐tolerant ventricular cells, which are contracting after electrical stimulation.…”