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Primary culture method for porcine liver cells

A technology of primary culture and hepatocytes, which is applied in the field of primary culture of pig hepatocytes, can solve the problems of small number of hepatocytes, complicated separation operation, easy contamination of hepatocytes, etc., and achieves small cell loss, simple operation, and less pollution Effect

Inactive Publication Date: 2017-09-12
YUNNAN AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a primary culture method of pig hepatocytes, which solves the problem of the small number and low activity of hepatocytes obtained by the direct separation and culture method in the prior art, the in situ "Seglen two-step collagenase perfusion method" and the half In situ perfusion separation is complex, costly, and easy to contaminate hepatocytes

Method used

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  • Primary culture method for porcine liver cells
  • Primary culture method for porcine liver cells

Examples

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Embodiment 1

[0032] This embodiment provides a method for primary culture of pig hepatocytes, which includes obtaining liver, liver tissue briquetting, liver cell culture, and primary cell identification in sequence, wherein the liver tissue briquetting is obtained by placing the liver in In a sterilized petri dish, cut it into pieces and place it in a 30-60 mesh sieve for rolling, and collect the filtered liver tissue into a briquette in a sterilized container.

Embodiment 2

[0034] This example is based on Example 1, and is further defined: the liver is from a newborn piglet, and the age of the newborn piglet is less than 5 hours, and the age of the pig refers to the time from birth to harvesting of the pig's liver.

Embodiment 3

[0036] This example is based on Example 1 and further defines: the acquisition of the liver is to sterilize the newborn piglets by immersing them in alcohol, then kill them by the neck, move them to an ultra-clean workbench, take out the liver, and strip off the fiber components , the fibrous components include capsule and blood vessels.

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Abstract

The invention discloses a method for primary culture of pig hepatocytes, which is characterized in that it includes sequentially obtaining the liver, briquetting liver tissue, culturing liver cells, and identifying primary cells, wherein the briquetting of liver tissue is obtained by The liver was placed in a sterilized petri dish, cut into pieces, placed in a 30-60 mesh sieve and rolled, and the filtered liver tissue pieces were collected in a sterilized container. The present application adopts the method of culturing tissue blocks by rolling them with mesh sieves, and obtains suitable pig liver tissue blocks by rolling them with mesh sieves, the size of which is suitable for adherent growth, and the growth of liver cells is good.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a primary culture method of pig hepatocytes. Background technique [0002] As an in vitro culture model, the isolation and culture of pig hepatocytes has important biological significance for the basic research of pig nutrition, biochemical substance metabolism and liver biological function research. The in vitro experiments of pig primary hepatocytes have good reproducibility, basically maintaining the original differentiation state and metabolic function of hepatocytes. The primary cells of pigs are consistent with the in vivo conditions, and can study the nutritional effects and regulatory mechanisms of pigs in the closest in vivo state. So far, there is no mature method for primary culture of pig hepatocytes for reference. Therefore, the isolation and culture of pig liver primary cells in this experiment can provide liver cell materials for scientific research on the role of liver i...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/067
Inventor 郭荣富李美荃张春勇陈克嶙安清聪
Owner YUNNAN AGRICULTURAL UNIVERSITY
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