Adult rat cardiac muscle cell separation method

A cardiomyocyte and separation method technology, applied in cell dissociation methods, animal cells, vertebrate cells, etc., can solve the problems of difficulty in culturing adult rat cardiomyocytes, and achieve the effects of high yield, long survival time and high activity

Inactive Publication Date: 2018-11-02
JIANGYIN CHI SCI
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Problems solved by technology

Most of the cells used in research come from the cultured heart of neonatal rats. Cardiomyocytes of neonatal rats and adult rats have great differences in shape, structure and function. As a biological experimental tool for cardiovascular research, adul

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  • Adult rat cardiac muscle cell separation method
  • Adult rat cardiac muscle cell separation method
  • Adult rat cardiac muscle cell separation method

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Embodiment Construction

[0022] In order to present the purpose and advantages of the present invention more clearly, specific embodiments will now be further described. The specific embodiments described here are only for explaining the present invention, and are not intended to limit the present invention.

[0023] The experimental equipment used in this experiment is as follows:

[0024] Inverted microscope (XDS-1A, Shanghai), ultra-clean bench (model: SW-CJ-1FD, Suzhou ultra-clean); cell culture box (model: 240i Thermo), super constant temperature water bath (model: HH-501 Shanghai Chuangyi Science and Education Equipment Co., Ltd.), constant flow pump (Longer Pump, BT 100-1L), langendorff perfusion system (Beijing Zhongshi Dichuang Technology Development Co., Ltd.).

[0025] In the present invention, SD rats with a body weight of 150-300 g are selected to be fed for 6-8 weeks, and cardiomyocytes of adult rats are isolated. The specific operation is as follows:

[0026] 1. Preheat the perfusion...

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Abstract

The invention provides an adult rat cardiac muscle cell separation method which comprises the steps: (a) preheating a perfusion system and fully cleaning a perfusion pipeline; (b) sequentially injecting heparin sodium and pentobarbital sodium into the abdominal cavity of a rat; (c) quickly opening the chest of the rat to expose the heart, taking the lower heart from the aortic root, putting into aperfusion solution, appropriately cleaning surrounding tissues and ligaturing the aorta on a perfusion needle; (d) connecting the intubated heart with the perfusion system and perfusing the heart bya perfusion solution without Ca<2+>; (e) continuously perfusing the heart by a mixed digestive enzyme solution; (f) taking the perfused heart, putting into a transfer solution, removing the atrium andgreat vessels, shearing the ventricle into small pieces, utilizing a suction pipe to blow and beat tissues into monodispersion cells, utilizing a filter screen to filter cell suspension and standing;(g) restoring Ca<2+> in a gradient mode; (h) precipitating cells after resuspending a complete medium to restore Ca<2+> and inoculating into a coated cell culture bottle to be cultured. By means of the adult rat cardiac muscle cell separation method disclosed by the invention, adult rat cardiac muscle cells which have high quality, high yield and long survival time can be obtained.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for separating adult rat cardiomyocytes. Background technique [0002] Cardiomyocytes cannot proliferate, and obtaining normal single cardiomyocytes is the basis of myocardial electrophysiological research. Most of the cells used in research come from the cultured heart of neonatal rats. Cardiomyocytes of neonatal rats are very different from adult rat cardiomyocytes in shape, structure and function. As a biological experimental tool for cardiovascular research, adult cardiomyocytes The functional characteristics of cells are irreplaceable for cardiomyocytes at other developmental stages, and an in vitro culture model of mature cardiomyocytes is necessary, but it is difficult to culture adult cardiomyocytes. The method for separating adult rat cardiomyocytes provided by the present invention can obtain adult rat cardiomyocytes with high quality, high yield and lo...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0657C12N2509/00
Inventor 蒋敏齐来俊其他发明人请求不公开姓名
Owner JIANGYIN CHI SCI
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