RU2538701C1 - Method for separating chicken embryo from egg for purposes of producing cell transplants from foetal tissues - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method for the non-traumatic separation of a chicken embryo consisting in taking a chicken egg with a 7-10-day embryo; the separation of the chicken embryo is preceded by the conservative trepanation of an egg shell by drilling 6 perforations within a blunt end and sawing egg shell spaces between the perforations from the inside, and removing the formed egg shell segment and the embryo.

EFFECT: method enables a nearly nine-fold increase of the separation ratio of undamaged chicken embryos from the egg.

1 ex

Description

The technical solution relates to the field of medicine, biology and biomedical technologies, in particular to a method for extracting a chicken embryo from an egg to further obtain cell transplants from its fetal tissues and can be used for the manufacture of biological products intended for the treatment of a wide range of diseases.

The well-known "Method for the preparation of a cell transplant of fetal tissues" [1]. In it, a cell transplant is obtained from fetal tissues of an abortive human fetus on weeks 17-21 of fetal development.

The disadvantages of this method are the moral, religious, legal contradictions that exist in the Russian Federation, the technical difficulties of directly collecting a cell transplant from fetal tissues of an abortive human fetus, as well as the high cost of this procedure, which undoubtedly inhibits the development of domestic transplantology. The way out of this situation may be to obtain cell transplants by extracting them from the fetal tissues of the chicken embryo. But, using the above-mentioned method, this is unrealistic, since the chicken embryo is surrounded by an egg shell and it is technically impossible to extract fetal tissues of the chicken embryo without its gentle trepanation.

A known method of preparing a cell transplant of fetal tissues of chicken embryos [2]. In it, to extract the chicken embryo from the egg, use the sharp end of the scissors (or a punch needle) to puncture the shell in the area of the pug, then cut part of the shell above the air chamber in a circle, trying not to touch the chorioallantoic membrane.

The disadvantage of this method is the high probability of trauma to the embryo when removing it from the egg.

The objective of the technical solution is to improve the method of extracting a chicken embryo from an egg to further obtain cell transplants from its fetal tissues, aimed at reducing the likelihood of injury.

The problem is solved due to the fact that a technique has been developed for extracting a chicken embryo from an egg, which reduces the likelihood of injury.

In the proposed method for extracting chicken embryo from an egg, the following difference from existing ones is provided, namely:

1. Refusal to puncture the shell with a needle or scissors and subsequent shearing of its segment with scissors.

2. Violation of the integrity of the egg shell by drilling 6 perforating holes in it, followed by sequentially sawing inside the intervals of the egg shell between the holes, followed by extraction of the obtained segment of the egg shell.

Between the set of essential features of the claimed object and the achieved technical result there is a causal relationship, which can be demonstrated by example 1.

Example 1

We conducted an experiment. 60 chicken embryos of 7-10 days of development were divided into two experimental groups of 30 each. In the first group, the extraction of embryos was carried out according to the method of Podchernyaeva R.Ya. et al., 2008, and in the second by our proposed method. In the first experimental group, 9 (29.9%) were damaged during the extraction from the shell, and in the second 1 (3.3%) embryos.

Our proposed method, “A method for extracting chicken embryo from an egg to further obtain cell transplants from its fetal tissues,” allows us to increase the percentage of extraction of intact chicken embryos from an egg by almost 9 times compared to the method used today.

We propose the following method for extracting a chicken embryo from an egg to further obtain cell transplants from its fetal tissues:

1. Take the chicken embryo 7-10 days of development.

2. Egg shells are treated with an antiseptic solution.

3. The gentle trepanation of the eggshell is carried out:

- Drilling in the area of the blunt end of the egg 6 perforating holes of the egg shell.

- Carrying out using a rigid conductor between the two nearest perforating holes of the elastic file.

- Gentle sawing with an elastic file from the inside of the egg shell gap between the pair of nearest perforating holes.

- We perform the same manipulations alternately with the rest of the spaces of the egg shell between the perforations.

- Removal of tweezers obtained by drilling and sawing a hexagonal segment of the egg shell.

- We get the embryo.

4. Select cells of various organs.

5. Suspend the selected cells of various organs.

6. We select suspensions with a high content of regional stem cells.

7. Using flow cytometry, we determine the number of stem cells in the selected suspension.

8. We subject the selected suspension of cryopreservation.

The technical and economic efficiency of the proposed method for extracting chicken embryo from an egg for further obtaining cell transplants from its fetal tissues is due to an almost 9-fold increase in the percentage of extraction of intact chicken embryos from an egg, which will certainly significantly reduce the cost of the resulting cell transplant and increase its number.

Information sources

1. Sukhikh G.T. "Method for the preparation of a cell transplant of fetal tissues" Patent for invention RU (11) 2160112 (13) C1 of the Federal Service for Intellectual Property, Patents and Trademarks, dated December 10, 2000

2. Podchernyaeva R.Ya. et al. Guidelines for working with cell cultures // Moscow, 2008, paragraph 7 (Internet: http://cells.virology.ru/home/93.html).

Claims (1)

Method for atraumatic extraction of a chicken embryo from an egg, characterized in that a chicken egg with an embryo of 7-10 day development is taken, before removing the chicken embryo, sparing trepanation of the egg shell is carried out by drilling 6 perforating holes in the region of the blunt end of the egg, followed by alternately sawing from the inside of the egg the shell between the holes, followed by extraction of the obtained segment of the egg shell and the extraction of the embryo.