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Method for recording T type calcium channel current by separating and cultivating newborn rat cortical neural cells

A technology for neonatal rat cortex and nerve cells, applied in the biological field, can solve the problems of short growth time, poor cell adhesion, poor neuron purity, etc., and achieves prolonged growth time, reduced chemical and mechanical damage, and small cell damage. Effect

Active Publication Date: 2012-12-05
辉源生物科技(上海)有限公司
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Problems solved by technology

[0004] The traditional trypsin chemical digestion method to separate cells and maintain cell growth in serum-containing medium is simple and easy, but there are problems such as poor adhesion of inoculated cells, short growth time and poor purity of neurons.

Method used

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Embodiment Construction

[0025] The present invention separates and cultures the method for recording T-type calcium channel current of newborn rat cortical nerve cells, comprising the following steps:

[0026] 1. Isolation and culture of neonatal mouse cortical neurons

[0027] Select a number of newborn 24-hour SD rats, male or female, sterilize the whole body with 75% alcohol, decapitate, cut the scalp and skull in layers under aseptic conditions, use curved forceps to open the field of view of the brain area, and carefully take out the whole brain. The dissecting fluid was washed several times. Carefully separate the cortex, place it in an ice bath for dissection, carefully remove the tiny blood vessels, and fully shred the tissue; add the same volume of 0.25% trypsin, cover the bottle with tinfoil, digest with 5% carbon dioxide, and digest in a 37-degree Celsius incubator for 20 for about 10 minutes, shaking several times during the period; after fully digested, add a few drops of fetal bovine s...

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Abstract

The invention provides a method for recording T type calcium channel current by separating and cultivating newborn rat cortical neural cells, and stably cultivated newborn rat cortical neural cells have a normal T type calcium ion channel function. The method is suitable to research on the physiological and pharmacological characteristics of the rat cortical neural cells by a patch clamp technique. The experiment operation is easy to master, and the method for stably cultivating the separated newborn rat cortical neural cells is simple and efficient. The success rate, the stability and the repeatability of an experiment are increased apparently by means of stably cultivating the separated newborn rat cortical neural cells.

Description

technical field [0001] The invention relates to biotechnology, in particular to a method for isolating and culturing newborn mouse cortical nerve cells and recording T-type calcium channel current. Background technique [0002] T-type calcium ion channels mainly exist in heart, nerve tissue, kidney, smooth muscle, sperm and various endocrine organs. T-type calcium current exists in the heart Purkinje fibroblasts, sinoatrial node, and potential pacemaker cells, and contributes to the repolarization of pacemaker cells, the initiation of action potentials, the self-discipline of the sinoatrial node, and the action of pacemaker cells Play an important role in the potential plateau. At the same time, T-type calcium channels are also involved in muscle excitation-contraction coupling, endocrine gland excitation-secretion coupling and cell regulation. [0003] Studies have proved that T-type calcium channels are not only important drug targets for the treatment of renal hypertens...

Claims

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Application Information

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IPC IPC(8): G01N27/26
Inventor 谈学海林佩元陈景才范柳
Owner 辉源生物科技(上海)有限公司
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