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Separation and culture method of epidermal stem cells of Beijing ducks

An epidermal stem cell and culture method technology, applied in non-embryonic pluripotent stem cells, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of low cell activity and purity, single method, high mortality, and achieve cell passaging The effect of many times, improved cell purity, and wide application range

Inactive Publication Date: 2013-04-03
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For poultry cell preservation, there are high requirements for cell purity and activity. At present, there are many disadvantages in the adherent culture method, such as single method, high cost, low cell activity and purity, and death rate after cell cryopreservation and recovery. too high

Method used

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  • Separation and culture method of epidermal stem cells of Beijing ducks
  • Separation and culture method of epidermal stem cells of Beijing ducks

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Isolation and cultivation of Peking duck epidermal stem cells

[0087] (1) Material collection: After killing Peking duck ducklings, grasp the head and tail tightly, and make the skin tight, and pluck the hairs in the opposite direction. Gently rub to remove the small hairs. After the hairs are clean, cut the skin with ophthalmic scissors, then gently tear off the skin with ophthalmic tweezers and put them in PBS with 100IU / mL penicillin and streptomycin respectively, and bring them back as soon as possible sterile room;

[0088] (2) Washing and digestion: After soaking the skin sample in 75% alcohol for 30 seconds, wash it with PBS or 0.75% normal saline with 100 IU / mL penicillin and streptomycin respectively, and wash it 10 times for 5 minutes each time. Carefully separate the subcutaneous connective tissue and most of the dermis, cut into 0.2cm×1-2cm strips, and finally cut into 0.2×0.2cm 2 Blot up the PBS or 0.75% normal saline around the skin tissue, ad...

Embodiment 2

[0091] Example 2 Identification of Peking Duck Epidermal Stem Cells

[0092] The Peking duck epidermal stem cells cultured in Example 1 were identified, the methods and conclusions are as follows.

[0093] 1. Observation of cell morphology

[0094] Methods: In the process of cell culture in vitro, the cells were routinely inspected to observe the growth status of the cells, the color change of the culture medium and whether the cells were polluted.

[0095] Conclusion: if figure 1 (Peking Duck Epidermal Stem Cells) shows that when the cells grow well, their shape is typical paving stone-like, with small cell volume, high nuclear-to-cytoplasmic ratio, and few organelles. The cells are spherical, compact, and have strong refraction under the microscope. The cell body is translucent, which proves that the cell growth is healthy and vigorous.

[0096] 2. Determination of cell viability

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Abstract

The invention provides a separation and culture method of epidermal stem cells of Beijing ducks. The separation and culture method comprises the following steps of: preparation of materials, washing, digesting, primary culture and subculture. The epidermal stem cells of the Beijing ducks prepared by the method can provide seed cells for life science researches in tissue engineering, gene engineering, cell engineering, immunology, molecular biology and the like, can enrich and improve local varieties in agriculture and open up a novel way in the aspect of protection of the genetic diversity oflivestock and poultry, and can be taken as main raw materials for producing vaccines.

Description

technical field [0001] The invention relates to a method for isolating and culturing duck epidermal stem cells, in particular to using the dorsal skin of Peking ducks to separate, cultivate, subculture and identify. in the field of cell biology. Background technique [0002] With the development of science and technology and the continuous improvement of people's material living needs, animal husbandry production is rapidly changing and developing in the direction of high yield, high quality, stress resistance, disease resistance and intensification. In most developed countries, due to people's excessive pursuit of economic interests, the livestock and poultry bred in large numbers are only a few varieties and hybrids with high economic value. Landraces with poor strength are endangered or even extinct. Although relevant international organizations, government departments, experts and scholars have attached great importance to the preservation and management of livestock g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074C12R1/91
Inventor 马月辉关伟军侯玲玲吕春荣白春雨郑杨
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI