UA56007U - Method for cryopreserving human spermia - Google Patents

Abstract

A method for cryopreserving human spermia includes mixing an ejaculate with a glucose-yolk-citrate cryopreserving agent comprising the cryoprotecting agent glyceryl and stage-by-stage cooling up to -196 °C.

Description

The useful model is based on the task of incubation, the mobility was 56.4x2.1905, and creating such a method of sperm cryopreservation 52.251.695, respectively. The obtained data on human motility, in which, due to changing the conditions of incubation of spermatozoa after rewarming, indicate a high level and cooling, the possibility of sub-cryoprotection of cells after using this would increase the motility of spermatozoa after rewarming, as well as cryopreservation itself. reduce the duration of the cooling process. Example 2. For the experiment, 4 ml of eggs were taken

This task is solved by the fact that in the method of cryo-culture with a concentration of cells of 56x106/ml and movement- preservation of human sperms, which includes a flux of 5253.8. 1.2 ml of sperm cryopreservation with glucose-yolk-citrate sideboard containing glucose-yolk-citrate cryopreservative, which contains cryoprotectant gly-buffer with glycerol content of 28 95, was added to the sperm, and placed in terceryl, and staged cooling to -1967C, according to mostat at 37 "C for 5 minutes. a useful model, the sperm is mixed with cryocon- Polypropylene straws with a volume of 0.5 ml zarvant in a ratio of 1:4, glycerol is taken in filled with the sample and transferred to the chamber the desired concentration is 28 95, and cooling is carried out from 3270 degrees, starting the cooling process. It is heated to 4"C at a rate of 4"C/min, after 5 minutes it is cooled from 32"C to 4"C at a rate of 4"C /min. cooling kicks are continued to -87C at the same speed After 5 minutes, cooling was continued to - speed, then seeding is carried out and continued to 87C at a speed of 4"C/min. Seeding was not carried out. Cooling to -507C at a speed of 10"C/min, after prolonged cooling to -507C at the speed at which the samples are immersed in liquid nitrogen. 10"C/min, after which the samples were immersed in liquid

A change in the incubation conditions (cryocon-nitrogen ratio (-1967C). Straws and sperm were heated on a water sideboard, increasing the concentration of glycine at 37"C for 5 minutes. The mobility of sperol) and the cooling regime, which provides for sperm were determined after heating ejaculate and seeding, allows to increase the cryoprotection was 55,452,3 90, after an hour of incubation of the cells, and thus to increase the motility of the sperm- 50,124,3 96, during the last two hours of incubation ate 1.5 times. The process lasts only 23 minutes. 46,053 ,5 95, and 24,032,7 95, respectively (Table 1).

The method is explained by the following examples. example, it can be seen that the absence of the seeding stage

Example 1. For cryopreservation, eyavel was taken for cryodamage of cells and reduction of motility with normospermia in a volume of 5 ml, with concentration during three hours of incubation. sperm ratio 75x105/ml and mobility 57 44.6 95. Example 3. In the experiment, ejaculate with

1.5 ml of cryopreservative was added to the sperm, which reduced sperm motility indicators (glucose-yolk-citrate buffer containing sperm). The samples were cooled from 32"C to 4"C volume of glycerol 28 95, and placed in a thermostat at a speed of 4"C/min. After 5 minutes of cooling, 37"C for 5 minutes. Polypropylene straws were heated to -8°C at a rate of 4°C/min. After 0.5 ml was filled with the sample and transferred to this, seeding was carried out by touching the straws in the program freezer chamber, starting with melt tweezers cooled in liquid nitrogen, and the cooling process. Cooled from 32"C to 4"C, continued cooling to -507C at a rate of 4"C/min. After 5 minutes of cooling at 10"C/min, after which the samples were immersed in liquid continued to -8"C at a rate of 4" s/min After nitrogen (-1967С). The straws were heated in water, seeding was carried out by touching the straws in a meta-bath at 37"C for 5 minutes. The data on the motility of left tweezers cooled in liquid nitrogen and warmed spermatozoa are shown in Table 2, from which the cooling to -507C was continued at a speed but that the process of cryopreservation of the samples at a temperature of 10"C/min, after which the samples were immersed in liquid nospermia did not affect the dynamics of nitrogen mobility (-1967C). Straws were heated in a water bath. Thus, this method can be used in a bath at 37"C for 5 minutes. The mobility of spe- used for cryopreservation of sperm ejaculates after rewarming was 62.351.8 90, after 1 hour of incubation both in normal and asthenospermia hours 60.1543.2 bo , in 2 and 3 hours

Table 1.

Indicators of human sperm motility. 11111111 Chaspisliavidirivutod.7///://Г/| 0 | 2 1 | 2 sh2 | With the method, 90 pom, 9o

Table 2.

Indicators of human sperm motility in asthenospermia. 11111111 Chaspisliaodprivugtod.7//-/://Г/| 07777112

Sources of information 2. dogde Naiyiyak, NKaKkezp K. Zpapta, SpPegui 1. And ocize UUeidei, Sai! 5. Riip5. Stoosigmima! oh MUeyvieaa, AvpokK Adagma!. Stguo-rgezeguayop oi

Ppitap z5reptaygoa ygogep ip eidni ayegepi rybeg Ppitap 5reptayog2oa: sotraigizop oi TE5T-uoIkK biyeg zuvietv// uoitpai Apago! - 1987. - M 8. - R. 41-47. apa diusegoi // Ipi. U. Repii. - 2000. - M. 45, Mo. 1. - R. 38-42.

Computer typesetting by I. Skvortsov Signed Circulation 26 approx.

Ministry of Education and Science of Ukraine

State Department of Intellectual Property, str. Urytskogo, 45, Kyiv, MSP, 03680, Ukraine

SE "Ukrainian Institute of Industrial Property", str. Glazunova, 1, Kyiv - 42, 01601