SU1750679A1 - Device for removing oocytes from the isolated ovary - Google Patents

Abstract

The device relates to agricultural biology and animal husbandry, is used for the reproduction of animals. The device consists of an adapter to which needles 2 and 3 of the same length are rigidly attached. Needles 2 and 3 in the middle of their length are rigidly fixed by clamp 4 which ensures a stable position of the needles and the necessary clearance between them. The needle 2, the outer diameter of which is 0.3 mm, communicates with the channel of the syringe 5 and serves to supply the medium into the cavity of the follicle. 5 mm through the flexible catheter 6 serves to drain the wash liquid 7 from the follicle cavity to the receiver 8. The distance between the outer edges of the needles 2 and 3 is 1.5 mm. which allows the extraction of oocytes from follicles having a diameter of at least 2.0 mm. 1 il.

Description

The invention relates to the field of agricultural biology and animal husbandry and can be used to extract oocytes from isolated ovaries under laboratory conditions for the purpose of cultivation, fertilization and their use for subsequent transplantation.

The significance of these studies is due to the need to use a huge reserve of germ cells (oocig) from outstanding cows. Over the entire period of domestic use, no more than 7–8 descendants are obtained from such animals, while in the ovaries of each animal, about 200 thousand germ cells are contained, and this valuable genetic reserve is lost with the delivery of the animal to the mill. Nowadays, many laboratories in the world are conducting experiments on

using this reserve to obtain a sufficiently large number of embryos, and from transplanting the last to recipient females, to achieve a multiple increase in the number of descendants from cows valuable in genetic terms. Realization of this problem requires the development and improvement of new methods, including on the issue of oocytes from isolated ovaries

 Two devices are known for extracting oocytes from isolated ovaries. One of them is a block of parallel inserted razor blades, which are cut the surface of the ovary and then repeatedly re-dipping the ovary with incisions in the Petri dish with the medium to wash off the oocytes from the cut follicles. This device allows relatively quick retrieval of oocytes. The disadvantage of it is that

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simultaneously with the oocytes, the tissue fluid, containing a number of decomposition products, which negatively affect the quality of the extracted oocytes, is washed away from the incisions

The other device is a syringe with a fitted injection needle. Extraction of oocytes using this device is performed as follows. About 5 ml of the medium are drawn into the syringe, the end of the needle is inserted through the ovary stroma into the follicle cavity and the b-b suction-filling-suction is obtained by extracting the oocyte from the follicle. This device allows this procedure to be performed more aseptically and eliminates the possibility of tissue fluid getting into the medium with the extracted oocytes. The drawbacks of the device are the long-term extraction procedure due to the need for multiple aspiration, the possibility of rupture of the wall of the follicle during the delivery of fluid into its cavity, and the relatively low oocyte recovery rate.

The aim of the invention is to accelerate the process of extracting oocytes from an isolated ovary without violating aseptic rules, ensuring a high level of extraction of oocytes and eliminating the possibility of tissue fluid getting into the leaching medium.

The goal is achieved by a device that includes a syringe with a nozzle for puncturing the wall of the follicle, the distinguishing feature of which is the presence of two parallel needles of the same length, rigidly fixed to the adapter, a needle of smaller diameter for supplying the medium to the follicle, and a needle of larger diameter for diverting fluid from the follicle to the receiver.

New in the device is a nozzle with two parallel spaced needles. The needles from the blunt end are rigidly fixed to the adapter, and in the middle of their length an additional retainer is installed, which provides the necessary clearance between the needles. The needle channel of a smaller diameter through the adapter communicates with the syringe channel. The parallel arrangement of the needles creates a vortex flow of washing liquid in the cavity of the follicle, which cuts the oocyte from the leg fixing it. The larger internal section of the discharge needle limits the increase in pressure of the washing fluid in the follicle cavity and prevents the follicle wall from breaking.

Testing the proposed device in the experiment more than 500 follicles

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with a diameter of 2 mm and more showed the maximum possible efficiency (100% recoverability of oocytes).

The drawing shows the proposed device.

The device consists of an adapter 1, to which the needles 2 and 3 of the same length are rigidly fixed. Needles 2 and 3 in the middle of their length are rigidly fixed with a lock

4. ensuring a stable position of the needles and the necessary clearance between them Needle 2, the outer diameter of the needle is 0.3 mm, communicates with the syringe channel 5 and serves to supply the medium into the cavity of the needle needle

The SC with an outer diameter of 0.5 mm through a flexible catheter b serves to drain the washing liquid 7 from the follicle cavity to the receiver 8. The distance between the outer edges of the needles 2 and 3 is 1.5 mm. which allows

extract oocytes from follicles having a diameter of at least 2.0 mm.

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The device operates as follows

Before use, it is sterilized by boiling in a disassembled form, and after sterilization and assembly, the internal channels of the device are washed with a sterile medium.

0 A medium 7 is brought into the syringe 5 with a capacity of 10 m and an adapter 1 with parallelly connected needles 2 and 3 is mounted on the tip of the syringe. Receiver 8 is connected to the back end of the discharge needle 3

5 flexible catheter b Take an isolated ovary and fix it in a position convenient for manipulation. The assembled device punctured the ovarian stroma to the bottom of the follicle. After login

0 sharp ends of needles 2 and 3 into the follicle cavity by lightly pressing the stem of the syringe 5 are fed into the follicle medium 7 through the needle channel 2. The volume of medium sufficient to flush the oocyte from the follicle is 2 ml and is fed into

5 one reception. Under the influence of the resulting pressure, the liquid from the follicle cavity through the channel of the withdrawing needle 3 and the flexible catheter b enters the receiver 8, while in the follicle cavity itself during the feeding period

In the medium, a vortex is generated in the circulation of fluid, under the influence of which the oocyte easily breaks off from the foot fixing it. Due to the larger cross-section of the internal channel of the diverting needle 3 relative to the needle

5 2 eliminates the risk of rupture of the follicle wall when the medium is introduced into its cavity. In this order, other surface follicles are washed, and one syringe filling with medium is enough to handle 5 follicles. The nozzle with para-needle system can be connected to the syringe via a standard three-way valve, which will allow filling the syringe with medium without first removing the syringe. The simplicity of the design of the device contributes to the rapid acquisition of the skill to extract oocytes, and the time spent on processing one ovary does not exceed that when using a device to cut the surface of the ovary.

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Claims (1)

Claims An apparatus for extracting oocytes from an isolated ovary, comprising a syringe with a nozzle for puncturing the follicle wall, characterized in that, in order to increase the efficiency of operation, the nl-cage consists of two parallel needles of the same length, the needle in the follicle, has a diameter smaller than the needle for draining fluid from the cavity of the follicle.