Abstract
We have derived a cardiac muscle cell line, designated HL-1, from the AT-1 mouse atrial cardiomyocyte tumor lineage. HL-1 cells can be serially passaged, yet they maintain the ability to contract and retain differentiated cardiac morphological, biochemical, and electrophysiological properties. Ultrastructural characteristics typical of embryonic atrial cardiac muscle cells were found consistently in the cultured HL-1 cells. Reverse transcriptase-PCR-based analyses confirmed a pattern of gene expression similar to that of adult atrial myocytes, including expression of alpha-cardiac myosin heavy chain, alpha-cardiac actin, and connexin43. They also express the gene for atrial natriuretic factor. Immunohistochemical staining of the HL-1 cells indicated that the distribution of the cardiac-specific markers desmin, sarcomeric myosin, and atrial natriuretic factor was similar to that of cultured atrial cardiomyocytes. A delayed rectifier potassium current (IKr) was the most prominent outward current in HL-1 cells. The activating currents displayed inward rectification and deactivating current tails were voltage-dependent, saturated at >>+20 mV, and were highly sensitive to dofetilide (IC50 of 46.9 nM). Specific binding of [3H]dofetilide was saturable and fit a one-site binding isotherm with a Kd of 140 +/- 60 nM and a Bmax of 118 fmol per 10(5) cells. HL-1 cells represent a cardiac myocyte cell line that can be repeatedly passaged and yet maintain a cardiac-specific phenotype.
MeSH Terms
Animals
Atrial Function
Atrial Natriuretic Factor/biosynthesis
Cell Differentiation
Connexin 43/biosynthesis
Electric Conductivity
Female
Gene Expression
Heart Atria/cytology
Heart Neoplasms
Mice
Mice, Inbred C57BL
Muscle Proteins/biosynthesis
Myocardial Contraction
Myocardium/cytology
Phenethylamines/metabolism
Phenotype
Sulfonamides/metabolism
Tumor Cells, Cultured
Chemicals
Connexin 43
Muscle Proteins
Phenethylamines
Sulfonamides
Atrial Natriuretic Factor
dofetilide
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Claycomb W C
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, New Orleans, LA 70112, USA. wclayc@lsumc.edu
Lanson N A
Stallworth B S
Egeland D B
Delcarpio J B
Bahinski A
Izzo N J
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