RU2428997C1 - Method of obtaining medications from fetus umbilical cord - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method of obtaining medications from fetus umbilical cord lies in the following: from calf fetus in the first part of intrauterine development umbilical cord as amputated in area of umbilical ring and on border of amnion-chorionic part of fetus placenta, blood is discharged from umbilical veins and arteries, amputated end of umbilical cord are fixed, umbilical blood is separated, components are separated, passed through nano-filters, amputated umbilical cord is disinfected and Wharton's jelly is separated, umbilical arteries, veins, umbilical envelope are separated and fixed at specified conditions, obtained materials are frozen in liquid nitrogen.

EFFECT: method makes it possible to increase purity of obtained preparation without application of chemical reagents.

1 tbl, 3 dwg

Description

The invention relates to veterinary medicine, and in particular to methods for producing Warton jelly, cord blood, umbilical cord veins and arteries and umbilical cord embryo membranes for future biocontainers, for targeted drug delivery in the treatment of cows with genital diseases after pathological birth and diseases of newborn calves with diarrhea .

A known method of obtaining nucleated cells from umbilical cord blood. The essence of this method is the collection of umbilical cord blood and the subsequent isolation from the blood of nucleated cells, in their subsequent conservation [1]. The disadvantage of this method is the complexity and duration of obtaining and processing umbilical cord blood before cryopreservation and after thawing.

A known method of obtaining a transplant isotolaryngology, where when creating typolastic material receive and use Wharton jelly. In the method, a portion of the umbilical cord of the fetus is resected, squeezed from blood clots from the umbilical vein and artery. After longitudinal dissection of the umbilical cord and separation of Warton's jelly, it is placed on a nylon mesh, such a transparent film, 0.5 ml thick, is applied to the wound [2].

The disadvantage of this method is that during the preparation of the Warton jelly it is laid in separate strands on a nylon mesh, exposed to ultraviolet radiation, and irrigated with a 1% dioxidine solution, which reduces the quality of the resulting embryonic material. As the closest analogue, a method for producing embroblast-like cells from the umbilical cord of a newborn is adopted [3]. The method consists in the fact that fibroblast-like cells are obtained from the umbilical cord of a newborn baby, when a fragment of the umbilical cord is exposed to collagenase-1 and collagenase-4 in a ratio of 1: 1 in the ratio of homogenate to solution 1: 5, removing endothelial cells from the umbilical cord vein.

The disadvantage of this invention is that to obtain natural therapeutic substances, the authors use enzymatic effects created by a chemical method, which reduces the healing properties of the obtained extracts, changing the structure of placental enzymes, and reduce the differential properties of natural embryonic cells.

The objective of the invention is to provide a method of obtaining from the umbilical cord of a fetal calf at the age of the first half of the intrauterine development of not only cord blood, but also of Varton jelly, umbilical cord veins and arteries as nanotube containers, embryonic-fetal porous shell of the umbilical cord, which allows to prepare and preserve as much transplant possible number of stem cells, nano-enzymatic placental fractions, increase reliability and guarantee the preservation of their viability during targeted delivery arstva to the sick body.

The feasibility of using stem cells, cord blood and enzymatic preparations of the fetoplacental complex are intended for the treatment of cows with diseases of the genital organs after pathological birth. The high proliferative potential of stem cells, the content of growth factors in preparations that stimulate cell division and bactericidal activity make them unique and indispensable in the treatment of animals.

The use of biological preparations includes mechanisms of physiological regeneration and inflammatory proliferation in the area of burn and stab wounds of the mammary gland in cows. The preparations made by us in liquid and ointment form, in which there are from 10 to 30% fractions of cord blood or Wharton jelly, stimulate the proliferation of granulosa cells and fibroblasts. The presence of stem cells in umbilical cord vein or arterial biocontainers, as well as in the umbilical cord amniochorial membrane introduced into the diseased organism, exhibit migration and proliferation in inflammatory tissues in the genital organs of the cow or in the gastrointestinal tract of newborn calves.

A large number of undifferentiated stem cells of a nanoregulatory nature of enzymatic preparations that effectively fulfill a therapeutic role are concentrated in the umbilical cord of calf fruits located in the uterus of a pregnant cow in the first half of pregnancy.

The method of obtaining medicinal preparations from the umbilical cord that we offer includes amputation of the umbilical cord in the region of the umbilical ring and at the border of the amniochorial part of the baby placenta, evacuating the umbilical cord blood from the umbilical veins and arteries, fixing the ends with a biological thread (catgut) of the amputated umbilical cord, blood is separated, red blood cells are separated, the plasma and leukocyte fractions are nanofiltered, the umbilical cord is placed in the heat chamber, the amputee is longitudinally dissected umbilical cord for free runoff of molten Wharton jelly, umbilical cord veins, arteries, umbilical casing are removed, cryopreservation is carried out.

The proposed method for producing drugs from the umbilical cord of the fetus is performed in the following sequence:

1. amputate the umbilical cord in the umbilical ring and on the border of the amniochorial part of the infant placenta in calf fetuses in the first half of fetal development;

2. fix the ends of the umbilical cord segments by dressing with biological thread (catgut) (figure 1);

3. make a blood sampling from the umbilical veins and arteries by vacuum (figure 2);

4. umbilical cord blood is separated, the erythrocyte fraction is separated, the fraction with plasma and nucleated cells of the neutrophilic group is passed through a nanofilter from 20 to 100 nm;

5. Disinfect the amputated umbilical cord by quartzization for 20 minutes;

6. place the umbilical cord in a thermal (35-40 ° C) chamber, hanging it in a curved state by the bandaged ends (figure 3);

7. make a longitudinal section of the wall of the umbilical cord, fixing it with a slit down to create a free run-off of the Varton jelly molten at a temperature of 40 ° C in a container (Fig. 3);

8. remove the umbilical casing having porous endothelial loose walls and fix it with a 0.9% sodium chloride solution;

9. the umbilical arteries are removed and their veins are fixed in a 09% sodium chloride solution;

10. prepare for freezing in liquid nitrogen the umbilical cord veins and arteries, the umbilical cord membrane, umbilical cord blood, Wharton jelly for long-term storage, to create bio-containers in the use of targeted drug delivery to the diseased animal organ.

Practical use

The use of components that make up the umbilical cord of the fruit, processed into treatment fractions within 2 hours after slaughter of pregnant cows on sick cows after calving, is shown in table 1.

Table 1 Therapeutic efficacy of drugs Healing fractions Cow diseases Cows (total) Of them recovered number % Cord Blood (30% ointment) Stab wounds of the thigh eleven 10 90.9 Biocontainer - Wharton Jelly Postpartum endometritis 12 10 88.3 Biocontainer (fractions: plasma, neutrophils, Wharton jelly) Purulent endometritis 9 7 77.7 The control Endometritis 12 8 66.6

From table 1 it is seen that after treatment of cows with stab-cut wounds of the thigh, lower legs with ointments with 30% fractionated umbilical cord blood daily for 7 days, recovery of the cows occurred after 10-11 days in 90.9% of the cows. Thanks to the treatment of cows with the help of biocontainers, which include Varton jelly, packed in an umbilical cord and inserted into the isthmic part of the uterine horn after 24 hours 3 times, 88.3% of the cows recovered after 12 days. After cows with purulent endometritis of a biocontainer (plasma, neutrophils, Wharton jelly) were introduced into the pelvic cavity of cows 3 times, recovery was achieved in 77.7% of cows. The recovery process was observed by the vivid manifestation of sexual hunting. In the control group, when the cows were treated according to the generally accepted method, using antibiotics, uterine drugs, recovery occurred in 66.6% of the cows 35-40 days after calving.

Literature

1. Isaev A.A., Mkheidze D.M., Grishina V.V., Prikhodko A.V. “A method for producing nucleated cells from cord blood”, RU No. 2343928 C1, 2007.04.03, application No. 2007112126/15.

2. Zabirov R.A., Kagan I.I., Rakhmatullin P.P. "Method for the production of tymplastic transplant", patent RU No. 2174016 C1, application No.2000115520 / 14, 06/14/2000.

3. Prikhodko A.V., Isaev A.A., Kiselev S.L., Lagarkova M.A., Kosheleva N.V., Saburina I.N., Melikhova B.C. Application for invention RU No. 2008111708 A, application No. 200811708/13, 13.13.2008.

Claims (1)

A method of obtaining medicinal preparations from the umbilical cord of the fetus, including the isolation of Warton jelly, umbilical cord blood, centrifugation and separation of blood into components, freezing of all obtained materials in liquid nitrogen, characterized in that in calves in the first half of intrauterine development, the umbilical cord is amputated in the umbilical region rings and on the border of the amnio-chorial part of the baby’s placenta, secrete blood from the umbilical veins and arteries, fix the amputated ends of the umbilical cord, separate the umbilical cord blood, separate the e portion passed through nanofilters, disinfect and amputated cord Wharton's jelly is isolated, recovered umbilical artery, vein and fix them in a 0.9% solution of sodium chloride, is recovered umbilical sheath and fixed in 0.9% sodium chloride solution.

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