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Method for separating rat hepatocytes

A separation method and liver cell technology, applied in the field of rat liver cell separation, can solve the problems of large consumption and expensive collagenase, and achieve the effects of low cost, high utilization of collagenase, and simple operation

Inactive Publication Date: 2015-05-20
XINXIANG MEDICAL UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the Seglen two-step perfusion method has been widely used at home and abroad, but this type of method still suffers from the high price and large consumption of collagenase, and the multicellular clumps obtained from the liver cell suspension

Method used

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  • Method for separating rat hepatocytes
  • Method for separating rat hepatocytes
  • Method for separating rat hepatocytes

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Embodiment Construction

[0021] In order to make the object, technical solution and advantages of the present invention clearer, the following examples further describe the present invention in detail. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0022] The invention takes SD rats, male, with a body mass of 200±20 g as objects, and separates rat liver cells. The specific operation is as follows:

[0023] Step 1, taking out the rat liver;

[0024] Rats were injected with heparin sodium, intraperitoneally anesthetized with pentobarbital sodium, disinfected with 75% ethanol, and opened the abdominal cavity; the intravenous indwelling needle was inserted into the portal vein and ligated, and the superior and inferior hepatic vena cava were ligated with surgical thread.

[0025] Step 2. Preheat the calcium-free perfusate at 35~38℃ and perfuse until the liver expands;

[0026] The perfusion flow ...

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Abstract

The invention provides a method for separating rat hepatocytes. The method comprises the following steps: (a) taking rat liver out; (b) perfusing the liver with calcium-free perfusate pre-heated to 35-38 DEG C until the liver is swollen; (c) cutting infrahepatic vena cava so that the blood flows until the liver becomes earthy yellow; (d) perfusing the liver with calcium-containing perfusate pre-heated to 35-38 DEG C for 3-7 minutes and (e) perfusing the liver with perfusate pre-heated to 35-38 DEG C and containing 0.05% IV-type collagenase for 1-3 minutes. The method for separating rat hepatocytes provided by the invention has the advantages of being simple in operation, small in usage amount of collagenase and high in hepatocyte yield and survival rate.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for separating rat liver cells. Background technique [0002] Hepatocytes not only play an important role in basic research, but also are widely used in medical related fields. Hepatocyte transplantation is also an effective method for the treatment of acute liver failure, liver cirrhosis, genetic metabolic liver disease and as a transitional treatment for waiting for orthotopic liver transplantation, so the quality of hepatocyte suspension is very important. How to obtain a large number of active hepatocytes through a relatively simple method has always been an important problem to be solved in basic research and clinical research. For a long time, the Seglen two-step perfusion method has been commonly used at home and abroad, but this type of method still has collagenase which is expensive, consumes a lot, and obtains multicellular clumps of liver cell suspension. Conten...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 邢雪琨陈红丽樊晋宇武红艳郏亚静
Owner XINXIANG MEDICAL UNIV
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