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Culture medium for enhancing ectogenesis of ox preantral follicle and application thereof

A technology of preantral follicles and in vitro development, applied in the field of animal husbandry, can solve the problems of not establishing bovine preantral follicles, etc., and achieve the effect of facilitating promotion

Inactive Publication Date: 2011-11-23
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, a set of system for in vitro culture of bovine preantral follicles has not been established.

Method used

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  • Culture medium for enhancing ectogenesis of ox preantral follicle and application thereof
  • Culture medium for enhancing ectogenesis of ox preantral follicle and application thereof
  • Culture medium for enhancing ectogenesis of ox preantral follicle and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, the acquisition of new culture system

[0024] 1. Acquisition of preantral follicles: Place the isolated bovine ovary obtained from the slaughterhouse in a glass plate with a diameter of 60mm, remove the excess mesangium and corpus luteum, then cut the ovary into two along the hilum, remove the medulla, and culture After washing twice in culture medium (M199 medium, GIBCO, catalog number: 31100-035), the follicles were cut from the ovarian cortex in both horizontal and vertical directions with a knife, and the obtained follicles and medium were blown with a dropper, and then Blow and beat 80-100 times with a straw with a diameter of 1mm, filter through a 200μm mesh screen, then blow and blow 80-100 times with a straw with a diameter of 0.5mm, filter with a 100μm mesh screen, centrifuge at 1500r / min for 5min, remove the supernatant, and add the precipitate Dilute and suspend the separation liquid, check the follicles under a stereo microscope, count and mea...

Embodiment 2

[0047] Example 2. In vitro culture of preantral follicles in a new culture system

[0048] Follicles are divided into preantral follicles and antral follicles. Aneurysm refers to the development and growth of follicles from a preantral non-antral state to an antral state.

[0049] method 1:

[0050] Step 1 of Example 1 to obtain preantral follicles in the medium 1 of Example 1, 39 ° C, 5% CO 2 , cultivated for 21 days. The control culture medium of Example 1 was used as a control.

[0051] The cavity-forming rate of the preantral follicles developed in vitro was counted. The experiment was repeated three times, and the results were averaged or average ± standard deviation.

[0052] The result is as follows:

[0053] After being cultured in medium 1 for 21 days, the average growth value of the diameter of the follicles was 167.8±35.7um, and the cavity formation rate was 34.38%;

[0054] The average increase in diameter of the follicles cultured in the control medium for 21...

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Abstract

The invention discloses a culture medium for enhancing ectogenesis of ox preantral follicle and application thereof. A method for preparing the culture medium for facilitating ectogenesis of isolated mammal preantral follicle comprises the following step of: mixing insulin, transferrin, sodium selenite, sodium pyruvate, glutamine, hypoxanthine, blood serum, follicle-stimulating hormone, luteotropin, estrogen, a PTEN (Pentaerythritol Tetranitrate) inhibitor with an alpha-MEN culture medium to obtain the culture medium. As proved by an experiment, ox preantral follicle is firstly separated from cortex ovarii, the obtained follicle is cultured in vitro in a culture medium and a control culture medium provided by the invention, and the diameter accretion and the cavitation rate of the follicle are observed. A result indicates that the cavitation rate and the diameter accretion value of the preantral follicle obtained by culturing with the culture medium provided by the invention are higher than those of the preantral follicle obtained with the control culture medium, and a necessary culturing system is provided for further excavation of a female reproductive resource.

Description

technical field [0001] The invention relates to animal husbandry (animal reproduction), in particular to a culture medium for improving bovine preantral follicle development in vitro and application thereof. Background technique [0002] There are tens of thousands of primordial follicles on the bovine ovary. During the process of growth and maturation, a considerable number of follicles degenerate and atresia, and only a small number of follicles develop to mature stage until ovulation. Among the female reproductive resources, oocytes are essential materials for the research and development of embryonic biotechnology such as in vitro fertilization, embryo transfer, sex control, embryo segmentation, animal cloning, and transgenics. The lack of sources of oocytes has restricted the progress of these technologies. Therefore, the development of ovarian preantral follicles is undoubtedly an effective way to solve this problem. [0003] Compared with common somatic cell culture ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 李向东李冬冬孙婧
Owner CHINA AGRI UNIV
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