Establishment of a human fetal cardiac myocyte cell line

Abstract

Human cardiac myocytes undergo degeneration, cytolysis, and necrosis in a number of clinical disease conditions such as myocarditis, dilated cardiomyopathy, and during episodes of cardiac allograft rejection. The precise cellular, biochemical, and molecular mechanisms that lead to such abnormalities in myocytes have been difficult to investigate because at present it is not possible to obtain and maintain viable cell cultures of human adult cardiac myocytes in vitro. However, human fetal cardiac myocytes are relatively easy to maintain and culture in vitro, but their limited availability and growth, variability from one preparation to another, and varying degrees of contamination with endothelial and epithelial cell types have made it difficult to obtain reliable data on the effect of cardiotropic viruses and cardiotoxic drugs on such myocytes. These thoughts prompted us to attempt to derive a cell line of human cardiac origin. Highly enriched human fetal cardiac myocytes were transfected with the plasmids pSV2Neo and pRSVTAg and gave rise to a cell line (W1) which has been maintained in culture for 1 yr. Morphologic and phenotypic analyses of W1 cells by flow microfluorometry and immunoperoxidase techniques indicate that the W1 cell line shares many properties of human fetal cardiac myocytes, but appears not to react with specific antibodies known to react with markers unique to human endothelial, epithelial, skeletal muscle, and dendritic cells. These preliminary data suggest that the W1 cells may provide a unique source of an established cell line that shares many properties ascribed to human cardiac myocytes.

Platelets Potentiate Brain Endothelial Alterations Induced byPlasmodium falciparum

“…Cells were incubated at 37°C in 5% CO 2 in water-saturated air. Primary HBEC-5i cells in their fifth passage from isolation were transfected using a plasmid designated pSVT, to produce a stable and immortalized cell line (20,74). This plasmid was a pBR322-based plasmid construct, containing sequences that coded for the large T transforming protein of simian virus 40 (SV40) and the Rous sarcoma virus long terminal repeat.…”

HL-1 cells: A cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte

“…Human fetal cardiac muscle cells transfected with SV40 T antigen can be maintained for over a year in culture (9). These cells expressed several markers of early fetal human cardiac myocytes but did not contract (9). Two other reports (8, 10) also have established that the expression of SV40 T antigen in neonatal rat cardiomyocytes induces myocyte proliferation.…”

“…These cells retain the expression of several cardiac-specific genes but have no organized ultrastructure and do not contract (12). Human fetal cardiac muscle cells transfected with SV40 T antigen can be maintained for over a year in culture (9). These cells expressed several markers of early fetal human cardiac myocytes but did not contract (9).…”

“…Other attempts to immortalize cardiac myocytes with SV40 large T antigen have used a variety of cardiac promoters in transgenic animals (5-7) or have used retroviral transfection of primary cells (8)(9)(10). Still other approaches have made use of the protooncogenes v-Myc and v-H-Ras to transform cardiac myoblasts (11,12).…”

Cardiac Toxicity

“…Determination of cardiac-specific toxicity on the cellular, biochemical, or molecular level demands the use of either primary cardiomyocytes isolated from cardiac tissue or cell lines, which mostly resembles cardiomyocyte physiology [95,96]. In contrast to test systems for detection of ion channel-related cardiac toxicity, the use of artificial cell lines only transfected with cardiac ion channels for determination of cardiac cytotoxicity is not an option.…”