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Preparation method of a primary cell line of neonatal piglet intestinal villi epithelium

A technology of primary cells and newborn piglets, applied in the biological field, can solve problems such as deviation, error, and error of experimental results

Inactive Publication Date: 2020-03-31
YUNNAN ANIMAL SCI & VETERINARY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, when it is necessary to carry out experimental research on single, purified small intestinal villous epithelial cells, there will be a large deviation, error or even error in the experimental results due to the small amount of small intestinal villous epithelial cells and small intestinal tissue cells
In addition, with the existing pollution control technology, that is, the addition of double antibodies, the contamination of this culture can hardly be controlled

Method used

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  • Preparation method of a primary cell line of neonatal piglet intestinal villi epithelium
  • Preparation method of a primary cell line of neonatal piglet intestinal villi epithelium
  • Preparation method of a primary cell line of neonatal piglet intestinal villi epithelium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] A method for preparing a primary cell line of neonatal piglet small intestinal villous epithelium, comprising the steps of:

[0068] Step (1), take the small intestine of a newborn healthy piglet within 1 day of age, and cut the small intestine into a 5cm-long small intestine section, then cut the small intestine section along the axial direction to expose the mucosal surface, and set aside;

[0069] Step (2): Rinse the mucosal surface of the small intestine segment treated in step (1) with washing solution until the washing solution is clear and translucent, then cut off the small intestinal villi with ophthalmic scissors, and then add the small intestinal villi to the new washing solution, Each gram of small intestinal villi requires 15 mL of flushing solution, then centrifuges, discards the supernatant, and obtains small intestinal villus tissue mass;

[0070] Step (3), adding the small intestinal villi tissue mass obtained in step (2) to the trypsin digestion soluti...

Embodiment 2

[0092] A method for preparing a primary cell line of neonatal piglet small intestinal villous epithelium, comprising the steps of:

[0093] Step (1), take the small intestine of a newborn healthy piglet within one day of age, and cut the small intestine into a 10cm-long small intestine section, then cut the small intestine section along the axial direction to expose the mucosal surface, and set aside;

[0094] Step (2): Rinse the mucosal surface of the small intestine segment treated in step (1) with washing solution until the washing solution is clear and translucent, then cut off the small intestinal villi with ophthalmic scissors, and then add the small intestinal villi to the new washing solution, Each gram of small intestinal villi needs 20mL washing solution, then centrifuge, discard the supernatant, and obtain the small intestinal villus tissue mass;

[0095] Step (3), adding the small intestinal villi tissue mass obtained in step (2) to the trypsin digestion solution i...

Embodiment 3

[0117] A method for preparing a primary cell line of neonatal piglet small intestinal villous epithelium, comprising the steps of:

[0118] Step (1), take the small intestine of a newborn healthy piglet within 1 day of age, cut the small intestine into a small intestine section with a length of 8 cm, and then cut the small intestine section along the axial direction to expose the mucosal surface, and set aside;

[0119] Step (2): Rinse the mucosal surface of the small intestine segment treated in step (1) with washing solution until the washing solution is clear and translucent, then cut off the small intestinal villi with ophthalmic scissors, and then add the small intestinal villi to the new washing solution, Each gram of small intestinal villi requires 17mL of flushing solution, then centrifuges, discards the supernatant, and obtains small intestinal villus tissue mass;

[0120] Step (3), adding the small intestinal villi tissue mass obtained in step (2) to the trypsin dige...

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Abstract

The invention relates to a newborn piglet intestinal villus epithelium primary cell strain preparing method and belongs to the biotechnological field. With the small intestine section of a newborn piglet as the raw material, the quick, convenient and high-efficiency newborn piglet intestinal villus epithelium primary cell strain preparing method is provided. Passage of a prepared pig intestinal villus epithelium primary cell strain can be conducted for over 20-30 generations. By the adoption of the method, a large number of healthy, single and purified pig intestinal villus epithelium primary cells can be prepared within a short period of time to be used for experimental study in the fields such as cytobiology, preventive veterinary medicine, virology, immunology and animal nutrition or pig dysentery viral vaccine production and antigen preparation. Meanwhile, the healthy, single and purified pig intestinal villus epithelium primary cell strain is prepared for the first time, the problem that the single and purified pig intestinal villus epithelium primary cell strain is unavailable in the life science research field is solved, and the method has a positive effect.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for preparing a primary cell line of intestinal villus epithelium of newborn piglets. Background technique [0002] Although the existing cell culture technology has been developed for decades and has made great progress, the progress of in vitro culture technology for some special tissues and organs has stagnated. Up to now, a single, purified porcine small intestinal villi has not been successfully prepared. Epithelial primary cell lines, not to mention the cultivation of porcine intestinal villous epithelial cell lines. The existing porcine small intestinal villous epithelial cells are usually prepared by aseptically taking out the fetal pig small intestine, cutting it together with small intestinal villous epithelium and basal tissue (including small intestinal smooth muscle and connective tissue) with scissors, and then passing it through 0.025% conc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 姚俊高林李华春
Owner YUNNAN ANIMAL SCI & VETERINARY INST
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