Ion channels in a skeletal muscle cell line from a Duchenne muscular dystrophy patient
Ion channels in a skeletal muscle cell line from a Duchenne muscular dystrophy patient
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DOI：
10.1002/mus.880170909
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发表时间：
1994-09
期刊：
影响因子：
3.4
通讯作者：
R. Caviedes;P. Caviedes;J. Liberona;E. Jaimovich
中科院分区：
文献类型：
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作者：
R. Caviedes;P. Caviedes;J. Liberona;E. Jaimovich
如别处所述，使用来自UCHT 1甲状腺细胞系的条件培养基在体外建立细胞系（RCDMD），该细胞系来源于取自患有杜氏肌营养不良症（DMD）的7岁患者的肌肉活检组织（Biochim Biophys Acta 1992;1134：247-255）。与通过相同方法建立的其他细胞系不同，RCDMD细胞对转化高度难治，并且所得细胞系生长缓慢，倍增时间约为72 h。此外，细胞在超过20次传代和15次传代后继续生长。该细胞系的一些特征包括不与抗肌营养不良蛋白抗体反应，存在二氢吡啶PN 200 - 110（Kd）= 0.3 ± 0.05 nmol/L和Bmax = 1.06 ± 0.03 pmol/mg蛋白和α-银环蛇毒素（Kd = 1.02 ± 0.17 nmol/L和Bmax = 4.2 ± 0.37 pmol/mg蛋白）的受体。电压钳结构中的膜片钳细胞在高血清完全培养基中生长时缺乏离子电流，但它们可以通过血清剥夺和添加激素和微量元素诱导分化。在分化培养基中5天后，观察到非失活延迟整流钾电流。在第12天，观察到A型失活钾电流以及瞬时内向电流。在不存在钠电流和钾电流的条件下，非常快速的激活和快速失活钙电流是明显的。该细胞系为研究DMD的病理生理学细胞机制提供了可能性。John Wiley & Sons，Inc.
A cell Iine (RCDMD), derived from a muscle biopsy taken from a 7‐yearold patient with Duchenne muscular dystrophy (DMD), was established in vitro using conditioned media from the UCHT1 thyroid cell line as described elsewhere (Biochim Biophys Acta 1992;1134:247–255). Unlike other cell lines established by the same procedure, RCDMD cells were highly refractory to transformation and the resulting cell line grew slowly with a doubling time of approximately 72 h. Further, cells continue to grow after more than 20 doublings and 15 passages. Some of the characteristics of the cell line include lack of reaction with antidystrophin antibodies and the presence of receptors for the dihydropyridine PN200‐110 (Kd) = 0.3 ± 0.05 nmol/L and Bmax = 1.06 ± 0.03 pmol/mg protein and for α‐bungarotoxin (Kd = 1.02 ± 0.17 nmol/L and Bmax = 4.2 ± 0.37 pmol/mg protein). Patch clamped cells in the voltage clamp configuration lack ion currents when growing in complete medium with high serum, but they can be induced to differentiate by serum deprivation and addition of hormones and trace elements. After 5 days in differentiating medium, noninactivating, delayed rectifier potassium currents are seen. At day 12, A‐type, inactivating potassium currents as well as transient inward currents are seen. In conditions in which sodium and potassium currents are absent, a very fast activating and fast inactivating calcium current was evident. The cell line offers the possibility of studying cellular mechanisms in the pathophysoplogy of DMD. © 1994 John Wiley & Sons, Inc.