Claycomb W C, Lanson N A, Stallworth B S, Egeland D B, Delcarpio J B, Bahinski A, Izzo N J
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, New Orleans, LA 70112, USA.
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2979-84. doi: 10.1073/pnas.95.6.2979.
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.
我们从AT-1小鼠心房心肌细胞瘤系中获得了一种心肌细胞系，命名为HL-1。HL-1细胞能够连续传代，同时保持收缩能力，并保留分化的心脏形态、生化和电生理特性。在培养的HL-1细胞中始终发现具有胚胎心房心肌细胞典型的超微结构特征。基于逆转录酶-PCR的分析证实了一种与成年心房肌细胞相似的基因表达模式，包括α-心肌肌球蛋白重链、α-心肌肌动蛋白和连接蛋白43的表达。它们还表达心房钠尿肽基因。HL-1细胞的免疫组织化学染色表明，心脏特异性标志物结蛋白、肌节肌球蛋白和心房钠尿肽的分布与培养的心房心肌细胞相似。延迟整流钾电流（IKr）是HL-1细胞中最主要的外向电流。激活电流表现出内向整流，失活电流尾是电压依赖性的，在>>+20 mV时饱和，并且对多非利特高度敏感（IC50为46.9 nM）。[3H]多非利特的特异性结合是可饱和的，符合单点结合等温线，Kd为140±60 nM，Bmax为每10(5)个细胞118 fmol。HL-1细胞代表一种心肌细胞系，它可以反复传代，同时保持心脏特异性表型。
