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Simple, economic and efficient method for in-vitro maturity of porcine oocytes

An oocyte and in vitro maturation technology, applied in the field of cell biology, can solve the problems of unfavorable large-scale production, high cost of industrial mixed gas, and difficult transportation, and achieve good washing effect, easy preparation, and convenient portability

Inactive Publication Date: 2010-05-19
ANHUI AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a simple, economical and efficient method for in vitro maturation of pig oocytes, which solves the problem of the high cost of industrial mixed gases used in the current method for in vitro maturation of pig oocytes, difficulty in handling, and disadvantages in production. Mass production issues when working in the field or in the field

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0013] 1. Preparation of reagents and culture medium

[0014] (1) Preheat the egg wash

[0015] Preheat the egg washing solution in a 38°C incubator for use in oocyte selection.

[0016] (2) Balance oocyte in vitro maturation solution (IVM solution)

[0017] Add 400 μl of IVM solution to each well of the four-well plate to make it cover the bottom, then cover the surface of the liquid with 400 μl of paraffin oil (mineral oil), and place in 5% CO 2 , in an incubator with 95% air, equilibrate for at least 4 hours.

[0018] 2. Ovary Collection

[0019] Take the ovaries from the slaughterhouse, put them into a thermos cup filled with penicillin G and streptomycin sulfate and normal saline at 28-35°C, and transport them back to the laboratory within 2 hours.

[0020] 3. Oocyte Collection

[0021] When aspirating follicles, use a 10ml sterile syringe equipped with a 18-gauge needle to aspirate follicles 3 to 6 mm above the ovary, and slowly inject the extract into a 15ml conica...

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Abstract

The invention relates to a simple, economic and efficient method for the in-vitro maturity of porcine oocytes, which comprises the steps of collecting an ovary, collecting oocytes from the ovary, sieving the oocytes and maturing and culturing in vitro. The method is characterized in that the entire process of maturing and culturing the oocytes in vitro is carried out in a manual lung gas expiration environment, 40 of the sieved ovarian cumulus-oocyte complexes are in one hole on average and are transplanted into a four-hole plate which is transplanted into an inflation bag, a bag opening is sealed by a sealing machine, respiratory gas exchange is carried out by testing personnel, the gas of the lung is blown into the inflation bag by an air blowing needle, air in the inflation bag is exhausted completely, the inflation bag is filled with the gas of the lung, sealed, put into a constant temperature tank with the temperature of 38 DEG C and cultured, and maturing and culturing are carried out for 42-44h.

Description

technical field [0001] The invention belongs to the field of cell biology, and particularly relates to a simple, economical and efficient new method for in vitro maturation of pig oocytes. Background technique [0002] Chinese patent application 200810218336.X discloses "A Method for In Vitro Culture and Maturation of Pig Oocytes", which includes the step of collecting ovaries, the step of collecting oocytes from the ovary, the step of screening oocytes and the step of maturing and culturing. Before the mother cell collection step, the collected ovaries are stored in a container filled with physiological saline, and the temperature in the container is kept at 32-34°C; the egg washing liquid used in the oocyte screening step contains 1 to 10 , the step of collecting oocytes from the ovary, the step of screening oocytes and the step of mature culture, is characterized in that, before the step of collecting oocytes, the collected ovaries are stored in a container filled with ph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
Inventor 曹祖兵随刘才李运生章孝荣张运海
Owner ANHUI AGRICULTURAL UNIVERSITY
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