KR101189738B1 - Direct Embryo Collector - Google Patents

Abstract

The present invention relates to a fertilized egg direct egg phase (DEC), and more particularly to a fertilization mechanism for collecting the fertilized egg directly used in bovine fertilized egg transplantation from the overovulation induced cow.
When fertilizing eggs using a fertilized egg direct eggplant according to the present invention, it is possible to fertilize the fertilized egg simply and effectively because it can be egged in the manner of 1way. In addition, it is possible to reduce the amount of perfusion effectively with a small amount of perfusate, and reduce the time of extraction to 1/3 compared with the conventional 2way or 3way method. Therefore, the fertilized egg laying process using the fertilized egg direct egg laying machine of the present invention not only reduces the stress of the test livestock, but also does not need to continuously stimulate the uterine tip of the livestock, and does not need a specific correction facility for egg farming in a short time in a general farmhouse. It is thought that fertilized eggs can be fertilized.

Description

Egg Embryo Collector {Direct Embryo Collector}

The present invention relates to a fertilized egg direct egg phase (DEC), and more particularly to a fertilization mechanism for collecting the fertilized egg directly used in bovine fertilized egg transplantation from the overovulation induced cow.

Embryos transplanted in Canada in the early 1970s have made great progress in North America and Europe. Embryo transplantation has been widely used in cattle as a means of genetic improvement. That is, a multiovulation fertilized egg transplant (MOET;) that improves the ability of the group early by improving the ability of the vacant cows with excellent ability to ovulate, recovering fertilized eggs, transplanting them into low-capacity groups, and producing large quantities of calves with excellent capacity at once. Multiple Ovulation and Embryo Transfer) methods are used. Recently, immature follicles are repeatedly collected from oocytes with confirmed ancestry and capacity using an Ov (Ovum Pick Up) technique, and in vitro fertilization and culture of the collected eggs to produce in vitro fertilized eggs. Technology is being applied.

In Korea, bovine fertilized egg transplantation took place in the early 1980s, and in 1982, the first fertilized egg transplanted calf was born by the professor team, Gil-Sung Jeong, Konkuk University.In the 1980s, a lot of research was carried out in domestic fertilized egg transplantation to lay the groundwork for practical use. In 1993, a calf was produced by in vitro fertilization by Hwang Woo-suk, a team of professors at Seoul National University. In addition, Hwang Woo-suk's team produced a nuclear-transferred cloned calf in 1995 and somatic cloned calves in 1999, and Dr. Kyung-Kwang Lee, a biotechnology research institute, raised dogs producing lactoferrin-secreted transgenic cows in 1996.

Industrially, the most important factor in determining the feasibility of fertilized egg transplantation in the animal industry is securing a large number of fertilized eggs produced by females having good traits and good male germ cells and conception rate. However, because it is not easy to secure large quantities of excellent fertilized eggs, we have devised overovulation method to secure large ovaries, in vitro fertilization method using eggs obtained from ovaries of slaughtered livestock, ovarian egg ovary method (OPU) and in vitro fertilization egg method. It became. In order to effectively obtain a good quality fertilized egg, most of the egg fertilization method is used in vitro fertilization method of over-ovulation.

The fertilized egg grows with the growth of follicles in the ovary with the estrus cycle, and the ovulation process occurs when the follicle bursts in the estrus, and the ovulation process occurs in the oviductal larynx, and the ovulated egg passes through the fallopian tube. It is moved by ciliary movement in the fallopian tubes to the site of the fallopian tube. When the egg reaches the fallopian tube, the egg is fertilized with sperm to form a fertilized egg. The fertilized egg moves from the fallopian tubes to the uterine tip for 6-7 days, and when it reaches the uterine tip, the fertilized egg reaches the uterus with loss of embryo or early blastocyst. While the uterus is suspended from the fertilization date 13 days after implantation to the endometrial epithelial cells. Fertilized egg eggs are fertilized during the period of 7-8 days when the fertilized egg passes through the fallopian tube and reaches the tip of the uterus.

On the other hand, in the conventional chaeran technique, surgical and non-surgical procedures are used. Among them, the fertilized egg chaeran is performed using a 3way catheter and a 2way catheter. In surgical procedure, open the empty cow supplying the fertilized egg to expose the female genitalia, insert the tube into the fallopian tube bulge or bulge junction, and then flow the perfusion solution to insert the recovery tube at the junction of the fallopian tube and uterine tip. There is a method of recovering the fertilized egg flowed into the perfusion solution, and a method of recovering the fertilized egg by recovering the perfusion fluid from the lower uterus after inserting the tube into the uterine tip. This method requires a laparotomy during the procedure, so it takes more time and cost than the non-surgical procedure in the process. In addition, there are a lot of treatment tools necessary for fertilized egg eggs, difficult to operate alone, it has the disadvantage that it is difficult to repeatedly recover the fertilized egg in a single female. Due to this problem, most of the cattle chaeran is carried out by non-surgical methods at home and abroad.

In the non-surgical procedure, as described above, a fertilized egg chaotic method using a 2way and a 3way catheter is implemented. Among these, a fertilized egg chaotic method using a 3way catheter is performed based on the time of artificial fertilization of an overovulated blank cow through the superovulation method. The fertilized egg reached to the uterine tip at ~ 8 days of age is recovered by repeating the injection and recovery of perfusion fluid as shown in FIG. The necessary components in the fertilized egg chaeran method using a three-way catheter is as shown in Figure 1, the fertilized egg embryo process is shown in Figure 2, first remove the intrarectal feces, put the left hand on the rectum fingertips below the rectal membrane Use your senses to find the ovary and locate the uterus where ovulation has occurred. When the location of the uterus where ovulation has been confirmed is confirmed, the cervical canal is inserted through the induction of the left hand inserted into the rectum by inserting a stainless steel 3way polycatheter into the rectum and then placing the catheter in the ovulated uterine tip. Connect the air injection hose to the catheter end with a syringe to fix the catheter reached to the uterine tip, and when a proper amount of air is injected using a syringe, a swelling line is swollen and fixed to the uterine tip. After catheter fixation, as shown in FIG. 2, a perfusion solution, a syringe infusion tube and a recovery tube, and a chafing cup for collecting fertilized eggs are connected to the catheter, and a regulator is installed in the infusion tube and the recovery tube, respectively, and the tube inlet is closed. After the preparation for the egg collection, use a syringe connected to the infusion tube to suck the liquid contained in the perfusion solution pack, open the inlet to the infusion tube and inject the perfusion solution to the uterine tip using a syringe to prevent perfusion fluid backflow Close the inlet of the inlet regulator. Then, using the left hand inserted into the rectum, open the inlet of the regulator in the collection tube while massaging the uterine tip, and collect the perfusate in the egg cup while collecting the perfusate liquid in the uterine tip, and open the regulator under the egg cup. The fertilized egg collected by the filter in the chaya cup is collected. The chaeran method using this 3way has a lot of components as shown in Figure 1, the procedure is complicated as shown in Figure 2 is difficult to perform alone. Therefore, two or more persons are required in the procedure. In addition, since the catheter is made of metal, the inner wall of the uterus may be damaged by the catheter, as shown in FIG. 2, and the flow of perfusion fluid takes about 1 hour or more to reduce uterine damage. Since the correction is essential, stress is generated in the animals used for the eggs due to long-term correction and perfusion fluid.

The fertilized egg chaotic method using a 2way catheter, another method of non-invasive surgery, simplifies the process to compensate for the shortcomings of the fertilized egg method using the 3way catheter, and changes the catheter material to a rubber material to reduce the stimulation of the uterine wall. The catheter bends well in the uterus. The dichroic process is a three-way, two-way system in which the inlet and the outlet of the perfusate flow into one of the inlets, and the air inlet to control the balloon for catheter fixing. It is a way to ovulate the fertilized egg by adjusting two tubes. As shown in FIG. 4, when the intrauterine catheter is injected, the injector is coupled to the catheter and injected into the uterine tip, and after the appropriate amount of air is injected using a syringe into the air inlet, the catheter is fixed. Insert the tube that combines the infusion and recovery tube that connects the tube to the perfusate pack at the inlet of the injector, open the regulator inlet of the infusion tube connected to the perfusate pack, insert the perfusate to the uterine tip, and fill the regulator inlet. Close, open the regulator in the recovery tube to recover the perfusate with the egg cup, and recover the fertilized egg. In the same way as a 3way catheter, a 2way catheter also stimulates the fertilized egg by stimulating it with the left hand that inserted the uterus tip into the rectum. Although the 2way catheter has improved the problem of 3way, it still has problems with egg infusion because of its long injection tube and recovery tube, and many components, and it is necessary to put the hand on the hind limb and stimulate the uterine tip continuously during the process. Correction is necessary, and a nesting place with a calibration frame is also necessary.

Accordingly, the present inventors have made diligent research efforts to overcome the problems of the prior art, and when the 1-way fertilized egg direct egg laying device is devised, it is confirmed that the egg laying process is simple, and the time for collecting eggs can be shortened. It was completed.

Therefore, the main object of the present invention is to provide a fertilized egg direct egg laying apparatus that can directly fertilize the fertilized egg directly without the existing complicated chaeran process.

According to an aspect of the present invention, the present invention is formed in the vicinity of the drain and suction port 11, the drain and suction port 11 formed for drainage of perfusion fluid and suction of the fertilized egg at the front end of the body (10) 10 is fixed to the air at the same time to prevent the back flow of the perfusion liquid 12 formed through the air inlet passage 13, the connecting portion 14 formed so as to pass through the drainage and the suction port 11 in the rear end of the body (10) Catheter 01 consisting of; A chaotic injector (02) formed to be inserted into the catheter (01) through the connector (14); A syringe (03) configured to inject the initial perfusate into the catheter (01) and to recover the perfusate containing fertilized eggs; An adapter (04) formed between the syringe (03) and the connector (14) to direct the syringe (03) through the catheter (01); Consists of a fertilized egg direct egg phase, characterized in that to fertilize the fertilized egg in a 1-way manner.

Specific chaeran process through the fertilized egg direct egg phase of the present invention is as follows.

As shown in FIG. 6, first, a chaotic injector 02 is inserted through the connector 14 of the catheter 01 along the dotted line, and injected from the vulva of the female livestock to the uterine tip. Thereafter, the air is injected through the air injection passage 13 to inflate the balloon 12 in the catheter 01 so that the catheter 01 is fixed to the uterine tip. When the catheter 01 is fixed, the egg injector 02 is removed and an opposite portion 41 of the adapter 04 connected to the syringe 03 containing the perfusate is connected to the connecting portion 14 of the catheter 01. do. Thereafter, the perfusion solution injection and the perfusion solution containing the fertilized egg are recovered using the syringe 03. The fertilization process is terminated by the recovery of the perfusion solution containing the fertilized egg.

The fertilized egg direct eggplant according to the present invention in 1 way, excluded the chaeran cup and its connecting pipe which is necessary in the existing 2way or 3way method. When using the egg cups at the same time, it is necessary to connect the tube to the egg cups and another mechanism for fixing the egg cups is required, but the present invention is to recover the perfusion solution containing the fertilized egg through the fertilized egg directly egg harvester with a syringe After completing the, by recovering the fertilized egg in the egg cup in the laboratory, etc., the whole process can be easily carried out without additional equipment such as a correction frame.

In the fertilized egg direct egg phase of the present invention, the egg injector is formed to be inserted into the catheter is characterized in that the egg injector for the egg when the catheter is inserted into the uterus serves as a guide.

Injection of the intrauterine catheter during fertilization of the fertilized egg is inserted into the catheter for implantation into the catheter, it is injected to the tip of the uterus, and the air is injected through the air injection passage so that the catheter is fixed in the uterus.

In the fertilized egg direct egg phase device of the present invention, the material of the chaeran injector is characterized in that the stainless steel.

In addition, the material of the catheter is characterized in that selected from the group consisting of silicone, non-toxic rubber silicone rubber and natural rubber latex. Catheter made of the above material can minimize the stimulation of the inner wall of the uterus in the process of ovulation.

In the fertilized egg direct egg phase machine of the present invention, the syringe is characterized in that the injection of the perfusion solution and the recovery of the perfusion solution containing the fertilized egg at the same time.

The fertilized egg direct eggplant according to the present invention is a method of fertilizing the fertilized egg by pulling the fertilized egg together into the syringe by the pressure of sucking the perfusion fluid injected by pulling the syringe by hand. Conventionally, the perfusion solution is recovered by using the position difference, that is, through the recovery tube located at a position relatively lower than the position where the perfusion solution is injected, so that the process of recovering the fertilized egg takes a long time, However, the perfusion solution consumed has many disadvantages. On the other hand, the fertilized egg direct eggplant according to the present invention applies a force to the syringe by hand, the applied force is transmitted to the fertilized egg through the perfusate, and the fertilized egg is swept into the perfusion fluid pushed like a wave by the force to suck into the syringe By recovering the fertilized egg, the fertilized egg can be more efficiently harvested with a small amount of perfusion liquid in a short time.

In the fertilized egg direct egg phase machine of the present invention, the adapter (adaptor) is characterized in that the adapter 41 for bonding with the catheter for chaeran and the adapter 42 for bonding with the syringe. The adapter 41 for the coupling with the chaotic catheter has the same outer diameter as the inner diameter of the connector 14 of the catheter 01, and the adapter 42 for the coupling with the syringe 03 is the tip of the syringe 03. It has the same inner diameter as the negative outer diameter.

The adapter serves to maintain a constant distance between the bovine genital and syringe when fertilized eggs. In the process of fertilization, cows often excrete urine and feces. Sudden physiological phenomena prevent foreign substances from getting into the syringe and protect the egg from the kicking of cows. In addition, it is easy to fix and detach the catheter in the process of egg collection, and facilitate the use of the syringe during the egg collection.

As described above, when fertilizing eggs using the fertilized egg direct egg laying apparatus of the present invention, it is possible to fertilize the fertilized egg simply and effectively because it can be egged in the manner of 1way. In addition, it is possible to reduce the amount of perfusion effectively with a small amount of perfusate, and reduce the time of extraction to 1/3 compared with the conventional 2way or 3way method. Therefore, the fertilized egg egg laying process using the fertilized egg direct egg laying machine of the present invention not only reduces stress of the test livestock, but also does not need to continuously stimulate the uterine tip of the livestock, thereby reducing the stress received by the livestock, and does not require egg laying facilities. It is thought that eggs can be fertilized in a short time in a normal farm.

Figure 1 shows the components required for egg fertilization using a 3way catheter (1; perfusate, 2; Flushing Accessories, 3; egg cup, 4; 50ml syringe, 5; Flushing Tubings, 6; polycatheter (3way fole)) .
Figure 2 is a schematic of the fertilized egg chaeran process using a 3way catheter.
Figure 3 shows the components required for fertilized egg eggs using a 2-way catheter (1; perfusate, 2; egg cup, 3; Flushing Tubings, 4; egg injector, 5; polycatheter).
Figure 4 is a schematic of the fertilized egg chaeran process using a two-way catheter.
Figure 5 shows the combination of the components of the fertilized egg direct egg phase group according to the present invention.
6 is a schematic diagram showing the composition of the fertilized egg direct egg phase according to the present invention.
Figure 7 is a schematic diagram of the process of fertilization using a fertilized egg direct egg laying machine according to the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by these examples.

Example  One. How to get eggs  1 time egg In chalan  Compare expenses

In general, the 3-way and 2-way chaeran method and the 1-way chaeran method using a fertilized egg direct eggplant according to the present invention, it is shown in Table 1 by comparing the expenses required for one-time fertilized egg chaeran.

As shown in Table 1, using the fertilized egg direct egg laying machine of the present invention, since it is possible to fertilize the egg with a small amount of perfusion liquid, it is a very cost effective method.

[Table 1. Expenses for one time egg fertilization]

Example  2. How to get eggs  Efficiency comparison

In the commonly used 3way and 2way chaeran method and 1way chaeran method using a fertilized egg direct eggplant according to the present invention, the efficiency of the flushing solution, the treatment time and fertilized egg egg was compared and analyzed, the results are shown in Table 2 below.

In each method, the egg harvesting process was carried out using a three-way, two-way harvesting process generally known, and the egg-processing process using the fertilized egg direct egg laying apparatus of the present invention was performed as described above.

As a result, the efficiency was the highest when the egg was fertilized using the fertilized egg direct egg laying machine of the present invention (61.7%), the procedure time was also reduced to 1/3 compared to other methods. Therefore, when fertilizing the eggs using the fertilized egg direct egg laying machine of the present invention provides a method of egg fertilization egg which is very effective in terms of cost and efficiency.

[Table 2. Efficiency comparison according to chaeran method]

Claims (6)

Drainage and suction port 11 formed for discharging the perfusion fluid and suction of the fertilized egg at the tip of the body 10, formed in the vicinity of the drainage and suction port 11 to fix the body 10 and at the same time backflow of the perfusion fluid A catheter (01) consisting of a balloon (12) formed through the air injection passage (13), and a connector (14) formed through the drainage and suction ports (11) at the rear end of the body (10) to prevent the; A chaotic injector (02) formed to be inserted into the catheter (01) through the connector (14); A syringe (03) configured to inject the initial perfusate into the catheter (01) and to recover the perfusate containing fertilized eggs; An adapter (04) formed between the syringe (03) and the connector (14) to direct the syringe (03) through the catheter (01); The fertilized egg direct eggplant, characterized in that to fertilize the fertilized egg in a 1-way manner.
2. The fertilized egg direct eggplant apparatus according to claim 1, wherein the egg injector formed to be inserted into the catheter allows the egg injector to act as a guide when the catheter is inserted into the uterus.
The fertilized egg direct egg laying apparatus according to claim 1, wherein the material of the egg injector is made of stainless steel.
The method of claim 1, wherein the material of the catheter is fertilized egg direct egg laying machine, characterized in that selected from the group consisting of silicone, silicone rubber and latex.
According to claim 1, The syringe is fertilized egg direct egg phase characterized in that the injection of the perfusion solution and the recovery of the perfusion solution containing the fertilized egg at the same time.
The fertilized egg direct egg sheller according to claim 1, wherein the adapter comprises an adapter (41) for bonding with a catheter for harvesting and an adapter (42) for bonding with a syringe.

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