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Cryopreservation method of primary-generation mouse parenchymal liver cells

A technique of hepatocyte and cryopreservation method is applied in the field of cryopreservation of mouse primary hepatocytes, which can solve the problems of unsatisfactory long-term cryopreservation effect, high price of UW solution, etc. It is beneficial to adhere to the wall and has a good effect of activity

Inactive Publication Date: 2018-07-10
JIANGYIN CHI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, some people use UW solution to cryopreserve hepatic parenchymal cells, but UW solution is expensive and can only store hepatic parenchymal cells at low temperature for a short time, and the effect of long-term cryopreservation is not ideal

Method used

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  • Cryopreservation method of primary-generation mouse parenchymal liver cells
  • Cryopreservation method of primary-generation mouse parenchymal liver cells
  • Cryopreservation method of primary-generation mouse parenchymal liver cells

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Embodiment Construction

[0019] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0020] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0021] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0022] The experimental instruments and reagents used in this experiment are as follows:

[0023] A set of surgical instruments, CCK-8 cell counting box (Sigma, USA), inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifuge (TD24B-WS, Shanghai), ultra-low temperature refrigerator ( Zhongke Meiling), pipette gun (Eppendorf, USA), electronic analytical balance (Sartorius, USA), ultra-clean bench (HJ-CJ-1D, Shanghai), CO 2 Cell incub...

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Abstract

The invention provides a cryopreservation method of primary-generation mouse parenchymal liver cells. The method includes the steps of firstly, preparing mouse parenchymal liver cell cryopreservationliquid; secondly, using a two-step perfusion method to separate the mouse parenchymal liver cells; thirdly, using the cryopreservation liquid to resuspend the mouse parenchymal liver cells, and then adding the mouse parenchymal liver cells into a cryopreservation tube; fourthly, using a step-by-step temperature reduction method to cryopreserve the mouse parenchymal liver cells; fifthly, resuscitating the mouse parenchymal liver cells; sixthly, using a dye exclusion method to measure the survival rate of the mouse parenchymal liver cells; seventhly, determining enzyme index activity; eighthly,observing the morphology of the mouse parenchymal liver cells; ninthly, performing ELISA method detection; tenthly, performing RT-PCR detection. The cryopreservation method has the advantages that themethod is simple to operate, many cells can be obtained after cryopreservation resuscitating, high survival rate is achieved, and the method is an ideal primary-generation mouse parenchymal liver cell cryopreservation method and capable of providing reliable cell resources for experiments.

Description

technical field [0001] The invention belongs to the technical field of cell culture in modern biotechnology, and in particular relates to a freezing method for primary mouse liver parenchymal cells. Background technique [0002] Hepatic parenchymal cells are used as seed cells for cell transplantation to treat chronic liver failure and as target cells for drug screening. The preparation and cultivation of large-scale, highly viable hepatocytes is the most basic link in these studies. Therefore, the separation, culture and cryopreservation of hepatic parenchymal cells are attracting increasing attention and become a hot spot in current research. [0003] Hepatic parenchymal cells are very sensitive to external conditions and have poor tolerance to cryopreservation. Various chemical and physical damages caused by cryopreservation and recovery may affect the vitality and function of hepatocytes. The cryopreservation technology of hepatic parenchymal cells has been studied for ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0284
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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