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Method for separating and purifying chicken's spermospore

A technology for separation and purification of primary cells, which is used in the preservation of rare species, tissue and cell engineering, and transgenic fields.

Inactive Publication Date: 2008-02-06
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports on the separation and purification of chicken spermatogonial cells at home and abroad

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0015] 1. Determination of the separation method of chicken spermatogonia

[0016] Aseptically collect the testes on both sides of the 3-6 day-old cock after hatching, and place them in a preheated Ca-free 2+ , Mg 2+ In the phosphate-buffered solution (PBS), the epididymis, fat pad and albuginea of ​​the testis were removed with pointed forceps, and then the tissue was divided into 1mm 3 For the left and right small pieces, gently blow and beat in PBS for 1-2 minutes, and then remove the supernatant after standing still. The precipitated tissue pieces are digested by the following three methods:

[0017] Method 1: 1mg / ml collagenase+1.5mg / ml hyaluronidase / 0.25% trypsin two-step digestion

[0018] Method 2: 1mg / ml collagenase + 0.25% trypsin + 0.25% trypsin three-step digestion

[0019] Method 3: 1mg / ml collagenase + grinding + 0.25% trypsin two-step digestion

[0020] The average cell survival rates obtained after digestion of testicular tissue by method 1, method 2 and me...

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PUM

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Abstract

The method of separating and purifying chicken's spermatospore relates to trnasgene, rare species preservation, tissue and cell engineering, and other fields. The present invention includes first separating chicken's spermatospore and subsequent Percoll separation and adherent purification. The present invention provides proper chicken's spermatospore separating and purifying method by means of separating chicken's spermatospore from chicken's testis tissue, and Percoll gradient and adherent purification, spermatogenic epithelium cell suspension with great testis cell number and high survival rate of testis cell is prepared while ensuring elimination of other somatic cell components.

Description

technical field [0001] The invention relates to the fields of transgene, preservation of rare species, tissue and cell engineering and the like. technical background [0002] Spermatogonia are the mother cells of male germ cells. During the life of an animal, spermatogonia can self-renew and generate new stem cells to maintain a constant number of stem cells. On the other hand, they can proliferate and differentiate into various stem cells under the regulation of their own genes or external signals. Stage sperm cells until spermatozoa. It is the only cell in male adult stem cells that is closely related to the genetic background of the next generation. Because of these potential values, spermatogonia has increasingly become the object of attention of many transgenic scholars. However, the proportion of spermatogonia in the testicular cells of humans and animals is very small. According to some data, there are about 10 spermatogonia in each testis. 8 cells, of which about 2...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/076
Inventor 李碧春吴洪
Owner YANGZHOU UNIV
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