Ischemic myocardial injury in cultured heart cells: Preliminary observations on morphology and beating activity

Abstract

An in vitro model of myocardial ischemia has been established with primary monolayer cultures of neonatal rat heart cells. Ischemic conditions were simulated in vitro by subjecting the heart cell cultures to various levels of oxygen and glucose deprivation. After the ischemic treatments, cultures of beating muscle (M) cells were evaluated for functional and morphological changes. The experimental protocol consisted of treatment with 20% or 0% O2 and 1000, 500 or 0 mg glucose per 1 of medium for 4, 12 or 24 hr. Control cultures were treated with 20% O2 and 1000 mg glucose. The morphological alterations induced by the deficiency of O2 and glucose in the medium were the formation of pseudopodia and cytoplasmic vacuoles; increased cytoplasmic granulation; and the formation of abnormal cell shapes, such as long, spindly shaped M cells. There was a time-dependent decrease in beating activity as the M cells were exposed to longer durations of ischemic conditions. However, if the cultures were replenished with complete medium (1000 mg glucose) and 20% O2, the cells regained their ability to beat.

An IRF5 Decoy Peptide Reduces Myocardial Inflammation and Fibrosis and Improves Endothelial Cell Function in Tight-Skin Mice

“…This sequence was synthesized using standard FMOC-based solid phase techniques, end-capped with acetate and amide, and named IRF5S. To mimic phosphorylation aspartate (D) was substituted for serine (S) as was done previously to modulate eNOS activity [ 18 , 23 ]. The “phosphorylated” decoy peptide (EL D W D AD D IRLQI D NPD) was synthesized, end-capped and named IRF5D.…”

Experimental models of cardiac physiology and pathology

“…The NRCM model is also a useful tool to study ischemia/reperfusion (I/R) injury. In 1977, Acosta and Puckett first reported effects of hypoxic environment on in vitro cell culture (30).…”

Experimental Models of Cardiovascular Diseases

“…Mechanical stretch can be applied directly on cells to simulate cardiac volume overload associated myocardial stretch [9]. In addition, ischemia-reperfusion is replicated by hypoxia-reoxygenation [10] or oxygen-scavenging compound (e.g., Na 2 S 2 O 4 ) treatments [11]. These stimulations often lead to cellular responses that closely represent cardiomyocyte changes in vivo setting, such as hypertrophy, apoptosis, autophagy, and fetal gene expression, rendering them a reliable model of cardiac diseases.…”