Method for culturing pig hair papilla cells

A technology of papilla cells and culture methods, applied in cell dissociation methods, animal cells, culture processes, etc., can solve the problems of increasing the probability of bacterial contamination, difficulty in attaching dermal papilla cells, and difficulty in obtaining dermal papilla cells, etc., to achieve shortening The effect of cell passage time, shortening culture time, and improving adhesion efficiency

Active Publication Date: 2021-07-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the special anatomical position of the dermal papilla, obvious biological characteristics, and difficulty in adhering to the wall, it is difficult to obtain high-purity dermal papilla cells, which restricts the research progress of hair follicle biology.
[0006] The traditional separation method of dermal papilla cells is by microdissection, but this method is labor-intensive, easy to cause operator fatigue, increases the probability of bacterial contamination, and the number of cells obtained is small, and the isolated dermal papilla cells are covered by the substrate. Membrane-coated, difficult to adhere to the wall and slow to migrate out
The commonly used enzymatic digestion method can obtain dermal papilla cells in batches, but due to the difference in hair follicle cycle development pattern and skin thickness in different parts, there are significant differences in the methods of extracting and culturing dermal papilla cells in different parts of different species, and Different culture conditions will affect the passage time of dermal papilla cells
The currently established dermal papilla cell culture method is mainly aimed at hair-using animals, and the dermal papilla of hair-using animals is very different from that of humans, which is not suitable for the research needs of human hair follicle science. However, the dermal papilla cell culture method for pigs It has not really been established, and it is impossible to provide high-purity dermal papilla cells in batches for research. Therefore, it is urgent to provide a method for extracting and culturing high-purity dermal papilla cells suitable for pig ear tissue, so as to further analyze and study human hair follicles. disease lays the groundwork

Method used

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  • Method for culturing pig hair papilla cells
  • Method for culturing pig hair papilla cells
  • Method for culturing pig hair papilla cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Screening of Pig Dermal Papilla Cell Digestion and Isolation System

[0035] 1.1 Screening of enzymes

[0036] The present invention adopts dispase II, type I collagenase and trypsin and certain time gradient to carry out pre-examination, and sample type and used enzyme are as shown in table 1:

[0037] Table 1 Methods of extracting dermal papilla cells from different species and comparison of their effects on pigs

[0038]

[0039] The present invention first uses dispase II to digest pig ear skin, but finds that the hair follicle structure is incomplete (such as figure 1 shown), dermal papilla cells could not be extracted. The present invention adopts type I collagenase gradient test, and finds that the digestion time of 4-5h has a good hair follicle stripping effect, but after only using collagenase to pick the pig hair papilla, it is found that it is difficult to remove the adhesion around the hair papilla, so the present invention adds Digestion wi...

Embodiment 2

[0051] Embodiment 2 adopts the pig dermal papilla cell subculture experiment of isolation culture of the inventive method

[0052] Cut the skin tissue from the thinner part of the pig’s ear within half an hour after slaughter, place it in a 50mL centrifuge tube without curling it, and immerse it in physiological saline, and complete the isolation and culture within 8 hours;

[0053] The method for culturing dermal papilla cells specifically includes the following steps: the pig dermal papilla cells are taken from the above-mentioned ear sample, specifically including the following steps:

[0054] S1. Take the ear sample out of the normal saline, cut the skin of the ear sample into 1.5cm*4cm long strips, wash it in normal saline, scrape off the surface dirt, and then wash it in iodophor solution for 2 minutes; then put it in Wash in normal saline, and then wash in 75% alcohol solution for 2 minutes; wash the ear samples in normal saline again, then wash in PBS (purchased from G...

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Abstract

The invention relates to a method for culturing pig hair papilla cells. The pig hair papilla cells are taken from pig ear skin tissues, I-type collagenase and pancreatin are used for digestion and separation, and a fetal calf serum culture medium is used for adding rat tail collagen for cell culture. By adopting the method provided by the invention, the epidermis and dermis structures are easy to distinguish, the hair follicle structure is clear and easy to separate, the cell passage time is shortened by about 13 hours, and the adherence is firmer. According to the method provided by the invention, the characteristics of the pig hair papilla cells can be kept within seven generations, and the purity is high, so that the pig hair papilla cells with high purity and relatively good passage consistency can be obtained in batches.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for culturing pig dermal papilla cells. Background technique [0002] The hair follicle is the most important accessory organ of the skin, and its development involves the interaction of ectoderm and mesoderm. The cyclical regrowth of the hair follicle (periodic cycle of anagen, catagen, and telogen phases) is characteristic of leading to the growth of new hairs and the shedding of old hairs. When the periodic regeneration function of hair follicles is abnormal, it can lead to hair follicle diseases such as alopecia, hirsutism and hair shaft disease. With the development of society, people pay more and more attention to the hair that affects life and work, and the research on the periodic development of hair follicles becomes more and more important. [0003] At present, the effective solution to the problem of hair loss is hair transplantation. Since hair tran...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0627C12N2509/00C12N2500/84
Inventor 丁向东秦晋邹全蒋尧
Owner CHINA AGRI UNIV
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