Serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue

A serum-free culture medium and ovarian cortex technology, which is applied in the field of cell biology, can solve the problems that primordial follicles cannot be cultured in vitro for a long time, restrict industrial development, teaching and scientific research, etc., achieve clear system components, improve growth and development. The effect of survival time, simple preparation method

Inactive Publication Date: 2021-03-09
SHIHEZI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the existing above-mentioned deficiencies, the object of the present invention is to provide a serum-free culture solution for in vitro culture of sheep ovary cortex tissue, which solves the problem that primordial follicles in ovarian cortex tissue cannot be cultured in vitro for a long time, which greatly limits its industry. Development and Teaching and Research Issues

Method used

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  • Serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue
  • Serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue
  • Serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue

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Experimental program
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Embodiment

[0030] 1. Configure culture medium

[0031] Basal culture medium: add α-MEM and DMEM at a volume ratio of 4:1.

[0032] Serum-free culture medium: add BSA 5 μg / mL, sodium pyruvate 0.2 mM, glutamine 3 mM, hypoxanthine 3 mM, insulin 10 μg / mL, transferrin 6.25 μg / mL to the above basal culture medium in sequence , sodium selenite 6.25 ng / mL, V C 50 μg / mL, penicillin 100 IU / mL, streptomycin 100 IU / mL.

[0033] 2. Obtain ovarian cortical tissue and fragments

[0034] The ovary tissue samples of Xinjiang military reclamation fine-haired lambs were collected by surgery, stored in 0.9% saline supplemented with 200 IU / mL penicillin and 200 IU / mL streptomycin at 37.5°C, and sent back to the laboratory within 1 h, and then added with Rinse twice in normal saline with 100 IU / mL penicillin and 100 IU / mL streptomycin, once in alcohol and twice in normal saline, and then remove the surface fascia and appendages of the ovary under a stereomicroscope.

[0035] Then under a stereomicroscope,...

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Abstract

The invention discloses a serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue. The culture solution comprises a basic culture solution and an additive, wherein the basic culture medium is a mixed solution of alpha-MEM and DMEM; the additive comprises the following components: 3-10 micrograms / mL bovine serum albumin, 0.10-0.50mM sodium pyruvate, 1-5mM glutamine, 1-5mM hypoxanthine, 3-10 micrograms / mL insulin, 1-5 micrograms / mL transferrin, 2-8 nanograms / mL sodium selenite, 10-200 micrograms / mL VC, 50-200IU / mL penicillin and 50-200IU / mL streptomycin, and the pH valueis 6.9-7.5. Through the synergistic effect of the components, the proliferation speed of interstitial cells is inhibited, the proliferation of granular cells is promoted, the defect that the three-dimensional environment of original follicle development cannot be maintained due to tissue collapse caused by excessive proliferation of the interstitial cells in the culture process of the ovarian cortex tissue is overcome, the growth and development condition and survival time of original follicles in the ovarian cortex tissue in vitro are effectively improved, meanwhile, the influence of unknowncomponents in serum on research is avoided, and the serum-free culture solution has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a serum-free culture medium for in vitro culture of sheep ovary cortex tissue. Background technique [0002] As the reproductive basis of female fertility, the ovary contains a large number of follicles in different developmental stages, and is the reserve of female fertility. Under physiological conditions, more than 90% of follicles exist in the ovarian cortex in the form of dormant primordial follicles, and are recruited and activated batch after batch under the action of reproductive-related hormones and growth factors to enter the stage of developing follicles. Unfortunately, with the development of the individual, most of the follicles are lost due to degeneration, and less than 1% of the follicles will develop into mature eggs and finally fertilize to produce offspring, which is a great waste of female animal fertility. At present, the utilization of follicles in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/90C12N2501/998C12N2500/30C12N2500/32C12N2500/40C12N2500/25C12N2500/38C12N2509/10
Inventor 彭夏雨孙凤霞康立超王静魏勇胡广东陈磊蒋松祁凤华
Owner SHIHEZI UNIVERSITY
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