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Chicken small yellow follicular membrane cell in-vitro culture method for improving cell survival rate

A technology of in vitro culture of small yellow follicles, applied in the field of bioengineering, to achieve a high survival rate

Pending Publication Date: 2021-10-08
JIANGSU INST OF POULTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the current research reports, there is no report on the in vitro culture of chicken yolk theca cells. Therefore, establishing an effective separation, purification and culture method is a prerequisite for the application of chicken theca cells. How to efficiently separate the theca cells with high purity , and the price is low, easy to popularize, and it is of great significance to further study the characteristics of poultry germplasm resources and explore the regulation mechanism of follicle development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Methods Cultured small yellow chicken theca cell Example 1 A method of increasing cell viability of the embodiment

[0019] The first step, the culture medium is disposed

[0020] Was added 1wt% of double-antibody in DMEM basal medium, 10% chicken serum.

[0021] The second step, producing hen sterilized, taking small yellow follicles

[0022] Selecting a peak into the egg, and continuously laying hens, filling a large beaker of 2000ml of 75% ethanol, sacrificed hen immersed in a beaker, taken after 2-3 minutes with sterile the abdominal cavity was opened scalpel, cut with a scalpel ovaries removed entire sterilization, the small yellow follicles 6-8mm separated and placed in petri dishes sterilized.

[0023] A third step of peeling the connective tissue, tissue layer pruning

[0024] After stripping small yellow follicles with forceps elbow layer of connective tissue, the forceps reverse bend fixed small yellow follicles, with ophthalmic scissors to cut small yellow follic...

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PUM

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Abstract

The invention discloses a chicken small yellow follicle membrane cell in-vitro culture method for improving the cell survival rate, and relates to the technical field of biological engineering. The method specifically comprises the following steps: mixing a membrane layer of chicken small yellow follicles with collagenase, digesting, and filtering after digesting to obtain a cell suspension; centrifuging the obtained cell suspension, wherein the obtained precipitate is the chicken small yellow follicle cells; and carrying out mixed culture on the obtained chicken small yellow follicle cells and a cell culture solution to obtain a chicken small yellow follicle cell line. By utilizing the in-vitro culture method disclosed by the invention, a large number of chicken small yellow follicle cells can be obtained in a relatively short time, the survival rate is relatively high and reaches 95% or above, and the smooth proceeding of a subsequent test of the chicken small yellow follicle cells is ensured.

Description

Technical field [0001] The present invention relates to the field of biological engineering technology, particularly to a method for in vitro culture of chicken small yellow theca cells of an improved cell viability. Background technique [0002] Hen ovarian and follicular development is a complex physiological process highly coordinated, by various endocrine, autocrine and paracrine regulation. Sexual maturation in the ovary of the hen, there follicular various developmental stages, early stages of follicular development, body surrounding the oocyte cells, and combined together, the original development of granulosa cells, thereby primordial follicles started to form. When the primordial follicle is selected as the primary follicle development, mesenchymal cells differentiate into the cell membrane layer. Granulosa cells and membrane cells are separated acellular basement membrane component composition, effectively prevent the direct contact with blood and granulosa cells. White...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2509/00C12N2500/80
Inventor 李春苗赵振华贾雪波黄华云王钱保黄正洋黎寿丰吴兆林屠云洁沈海玉薛龙岗
Owner JIANGSU INST OF POULTRY SCI
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