Culture method of Jinhua pig mammary gland epithelial cell line

A technology of mammary epithelial cells and culture methods, which is applied in the field of culture of Jinhua pig mammary epithelial cell lines, can solve the problems of population decline and disappearance, and achieve the effects of stable quality, high cell purity, and reduced culture costs

Inactive Publication Date: 2015-09-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the negative consequences of improvement are that some landraces are gradually replaced by cultivated

Method used

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  • Culture method of Jinhua pig mammary gland epithelial cell line
  • Culture method of Jinhua pig mammary gland epithelial cell line
  • Culture method of Jinhua pig mammary gland epithelial cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Construction of Jinhua Pig Mammary Epithelial Cell Line

[0068] (1) Primary culture: Quickly cut mammary gland samples from Jinhua pig mammary glands that have been lactating for 21 days, wash with PBS plus double antibodies (penicillin and streptomycin) for 3~4 times, and then cut into 0.5~1.0mm 3 tissue block; move the tissue block into a 50ml centrifuge tube, add collagenase IV digestion solution to digest breast tissue for two hours, add an equal proportion of trypsin and collagenase IV mixed digestion solution to digest breast tissue for two hours, and filter the digestion solution in 100-mesh cell sieve, collect the filtrate, and transfer to a 15ml high-pressure sterilized centrifuge tube, centrifuge the cells at 1000rpm for 5min, pour off the digestion solution, keep the cell mass at the bottom, wash with PBS three times, add an appropriate volume of 10% fetal bovine Serum in DMEM culture medium, pipet the cell mass to make it evenly suspended, and fin...

Embodiment 2

[0076] Example 2 Detection of biological characteristics of Jinhua pig mammary gland cell line

[0077] The Jinhua pig mammary gland cell line that embodiment 1 cultivates is carried out the inspection of biological characteristic, and method is as follows:

[0078] 1. Cell growth status and color change of culture medium.

[0079] in conclusion:

[0080] Such as figure 1 It is Jinhua pig primary mammary gland cells, where A is mixed with fibroblasts and epithelial cells, and B is only epithelial cells after purification; figure 2 It is the micrograph of Jinhua porcine mammary gland epithelial cells after cryopreservation and recovery, where A is 24 hours after recovery, and B is 48 hours after recovery. Observed by an inverted microscope, the cells are polygonal and uniform in shape, and the cells are closely arranged to form a cobblestone-like monolayer aggregation, which has typical morphological characteristics of epithelial cells. After cryopreservation and recovery...

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Abstract

The invention discloses a culture method of a Jinhua pig mammary gland epithelial cell line. Jinhua pig lactation-period 21-day mammary tissues are subjected to primary culture and subculture to finally obtain the high-purity high-viability Jinhua pig mammary gland epithelial cell line. The mammary gland cell line belongs to the field of biology. The cultured mammary gland cell line is free of fibroblasts and other impure cells, and has high cell purity. The cryopreserved cells have stable quality. The viability of the cryopreserved and resuscitated cells can reach 90%. The cells grow stably after being subjected to subculture, and are suitable for large-scale culture. The cytological characteristic detection indicates that the cell line satisfies the cell line establishment requirement. The Jinhua pig mammary gland epithelial cells can be used for preparing a nucleus transplantation donor of a transgenic pig. The Jinhua pig mammary gland epithelial cell line can provide a raw material for research in medicine and molecular biology. The Jinhua pig mammary gland epithelial cell line can enrich and improve the local pig species in agriculture, and is an effective protection means for favorable pig species in China.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for cultivating Jinhua pig mammary gland epithelial cell lines. Background technique [0002] The issue of animal genetic resources protection was first raised in the United States in the 1950s. Since the Stockholm Conference on the Human Environment in 1972, it has attracted the attention of the Food and Agriculture Organization of the United Nations (FAO) and the Environment Program (UNEP). They have jointly carried out conservation actions for domestic animal genetic resources around the world, and have worked for decades. Obvious results have been achieved. Our country has participated in a series of plans and actions organized by them from the beginning. However, the current situation of China's livestock and poultry genetic resources protection is not optimistic. In the census conducted in the 1980s, it was estimated that there were 600 varieties, of which 385 were l...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 彭静徐宁迎
Owner ZHEJIANG UNIV
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