The method of obtaining material containing cells of the prostate gland, the material containing cells of the prostate gland, and methods of treatment of fibromatosis cancer, chronic prostatitis and disorders of the male reproductive function by transplanting

 

(57) Abstract:

The invention relates to medicine, namely to a method for producing a material containing cells of the prostate neonatal, young and adult rabbits, the material containing cells of the prostate obtained by the proposed method, and to transplantologists treatment of fibromatosis cancer, chronic prostatitis and disorders of the male reproductive function by transplantation of material containing cells of the prostate. The technical result related to the production of a material containing cells of the prostate gland, and to the actual material, is that the use of the prostate gland of a mammal, with the extracted prostate washed, cut it into slices and seeded received material culture mattresses, adding culture (growth) environment and serum mammals, then provide the optimum incubation. The technical result relating to methods of treatment of fibromatosis cancer, chronic prostatitis and disorders of the male reproductive function, is that transplantations are made of a material containing cells present the Ghana and tissues. 5 C. and 32 C.p. f-crystals.

The invention relates to medicine, namely to a method for producing a material containing cells of the prostate gland and intended for transplant patients, the actual material containing cells of the prostate obtained in a new way, and to transplantologists treatment of fibromatosis cancer, chronic prostatitis and disorders of the male reproductive function by transplantation of the received material.

A method of obtaining material from fetal human and animal (fruits pigs, fruits cattle), containing, in particular, islet cells of the pancreas. The method consists in the fact that the pancreas of fetal human, cattle or pigs extracted under sterile conditions, washed in Hanks solution with antibiotics, cut into fragments of a size of 2-3 mm, which again washed with Hanks solution with antibiotics. Then the fabric is dipped in a 0.1 to 0.3% solution collidine (preparation containing collagenase) or a mixture of equal parts of 0.1% solution collidine and 0.1% trypsin solution for 20-30 min at a temperature of 20-22. Then remove the solution fermentation 15-20 minutes Make two or three plum trypsin solution with suspended therein by single cells and small cell complexes in centrifuge bottles, and then centrifuged and the precipitate resuspended. Received not settled cells and cell complexes petroccione separated and sow their cultural mattresses. Incubation is carried out at a temperature of 36-37oC for 8-10 days with replacement of the growth medium every 3-4 days. When this was obtained, the culture, the bulk of which are beta-cells (see Experimental foundations of clinical islet cell transplant. /Shumakov Century. And., Blumkin C. N., Skaletsky H.N., The Shal'nev B. I. in the book "the Transplantation of islet cells of the pancreas (essays)", M, "Canon", 1995, S. 38-42).

The disadvantages of the known method is a significant loss of cells in the process of multi-material processing, in particular when the magnetic razmeshivali and by centrifugation, and fractional precipitation of cellular complexes and single cells.

Known material containing beta-cells derived above (see ibid.).

The disadvantages of the known material obtained in this way, it is what it is destroyed, as part injured while on a magnetic razmeshivali and by centrifugation.

A method of obtaining suspensions of viable cells, mammalian cells - interstitial cells of the testes of a human embryo, cows, pigs and newborn piglets and calves, and newborn animals should be as early age to not have time to emerge as a stable antigenic species specificity. For the preparation of suspensions used solutions containing nutrient substrate for cells, in particular using the compositions of the standard environments for cells animal tissue environments Needle, solution, Hanks, medium 199 and the like. In this case, all operations are carried out under aseptic conditions. The resulting suspension should be sterile (see the patent of Russian Federation N 2078590, A 61 M 37/00 A 61 K 35/48, 10.05.97).

There is a method of treatment of fibromatosis of the uterus with the use of androgens and anabolic steroids (see Trabanco So on, solski J. P. Handbook of obstetric-gynecological endocrinology". Kiev, Health, 1989, S. 196-197).

There is a method of treatment of fibromatosis of the uterus with the use of an agonist of gonadotropin-releasing hormone (see J. lane. and others Use the", N 2, 1991, S. 17).

However, these methods of treatment are not effective in some cases. In addition, a certain percentage of patients after treatment, resulting in decreased tumor size, subsequently, the tumor was again increased.

There is a method of treatment of chronic prostatitis, which consists in the fact that the patient is under the capsule of both lobes of the prostate impose a suspension of viable Leydig cells (cells embryonic testicular tissue) mammalian embryonic origin and newborns in the proportion of not less than 2 million cells in 1 ml (see the patent of Russian Federation N 2078590, A 61 M 37/00 A 61 K 35/48, 10.05.97).

There is a method of treatment of patients with impaired male reproductive function, namely, that the patient is under the protein shell of the egg is injected suspension of Leydig cells (cells embryonic testicular tissue) in an amount of not less than 2 million cells in 1 ml (see the patent of Russian Federation N 2026643, A 61 In 17/00 A 61 K 35/48, 20.01.95).

The disadvantages of these methods include some of their complexity and the risk of traumatic damage of the relevant authorities, with possible development in the subsequent inflammatory process (including autoimmune nature) of the prostate gland or under the tunica albuginea of the testis.

The invention related to a method for producing a material containing cells of the prostate gland, is its simplification, the increase in guaranteed high percentage (90%) extract of intact cells from the source material and the reduction due to the rejection of the use of the enzyme containing collagenase.

The invention related to a material containing cells of the prostate gland is the production of a material with high biological activity.

The invention related to methods of treatment of fibromatosis cancer, chronic prostatitis and disorders of the male reproductive function, is to increase the effectiveness of treatment through the use of a material with high biological activity.

The problem is solved in that when receiving material containing cells of the prostate gland and prostate young adult or newborn rabbit (prostate other mammal) with the extracted prostate washed in Hanks solution with the addition of antibiotics, cut it into slices and seeded received material cultural the initial (optimal) mode, for cultivated microfragments prostate gland, which carry out incubation at a temperature of 35 - 40oC continued from two days to six weeks with single or multiple temperature drops to 4 - 25oC and aged at a low temperature and with a temporary one-time or periodic decrease in the concentration of serum.

The task for the material containing cells of the prostate gland, is solved through the use of the above method thereof.

The task for a method of treatment of fibromatosis uterus is solved in its implementation, which produce a transplantation material containing cells of the prostate gland, while the above-mentioned material can be transplanted in various organs and tissues: subcutaneously, intramuscularly, in particular in the straight muscle of the abdomen, the liver directly into the parenchyma or via the portal vein or its branches), in the pulp of the spleen, splenic vein, in the peritoneal cavity, in a specially created muscular pocket in the liver parenchyma or in the greater omentum.

The principle difference of this method of obtaining material containing cells of the prostate gland, which, as source material used prostate newborns, young or adult rabbits, not embryos, as the nearest equivalent. For the proposed method is feasible and the use of embryos in mammals, but this is impractical because of the glandular structure in the embryonic prostate gland has not yet been formed.

When implementing the method in accordance with the present invention excludes the operation of traumatic cells of the prostate gland, namely the centrifugation, the processing on the magnetic razmeshivali, and the operation processing enzyme for dissolving the collagen fibers. In accordance with the present invention can provide a release of more than 90% of prostate cells contained in the original prostate gland.

To obtain a material containing cells of the prostate gland, can be used prostate mammals, in particular newborn mammals or their fetuses. Experience shows that the use of the prostate gland of cattle and their fetuses, as well as horses and pigs, fruit person, but most available and cheap material assetsother cells of the prostate gland, remove the prostate gland, such as young or adult rabbit, and put it in cold Hanks solution with the addition of antibiotics. Removing the prostate gland from the solution, remove the capsule and a layer of connective tissue with blood vessels. Decapsulated prostate gland is crushed to microfragments size up to 1 mm, and again washed with Hanks solution with the addition of antibiotics. The obtained material is seeded with a culture mattresses, which is added to the culture (growth) medium and serum of mammals. As serum can be used serum of cattle. Then sown cultural mattresses placed in thermostat, which ensures the optimum mode for prostate cells, which are incubated at a constant temperature in the range 35 - 40oC for from two days to six weeks, while momentarily reduce the temperature depending on the need (for culture is microscopic control) to 4 - 25oC and incubated for a certain time at a low temperature. The shutter speed is determined experimentally for a particular type of cell culture. For recip is the things or recurrent decrease in the concentration of serum. The exposure time at low serum concentration is also determined experimentally for a particular type of culture the cells of the prostate gland.

The resulting material can be used for transplantation immediately after collection or stored for quite a long time before the operation at low temperature or by freezing.

A method of treating fibromatosis of the uterus is performed with the use of material containing cells of the prostate obtained using the proposed method.

The choice of the method of introducing material containing cells of the prostate gland, may be different, i.e. the material can be transplanted in various organs and tissues: intramuscularly, in particular in the straight muscle of the abdomen, the liver (in the parenchyma or via the portal vein or its branches), in the pulp of the spleen, splenic artery, in the peritoneal cavity, in a specially created muscular pocket in the greater omentum or subcutaneously.

The simplest techniques are effective and least traumatic for the patient is the introduction of material containing cells of the prostate gland, by means of a syringe into the straight muscle of the abdomen.

The use of material containing cells of the prostate obtained by the proposed method for transplantation will significantly improve the efficiency of operations to simplify getting and to reduce the cost of the material obtained, which in turn is a prerequisite for the widespread implementation of the above methods of treatment.

Below described examples illustrate the effectiveness of the transplantation material containing cells of the prostate gland, when fibromatoses cancer, chronic prostatitis and disorders of male sexual function.

Example 1. Patient S. E., 52 years old, was admitted to the gynecological Department with a diagnosis of fibromatous of the uterus. When ultrasound on the left lateral wall of the uterus defined area of fibromatosis without distinct outlines a mean diameter of 23 mm of Myometrium multiple heterogeneous echostructure. Conclusion: fibromatous body of the uterus. Later, 4 days after admission the patient was performed intramuscular transplantation of material containing cells of the prostate obtained from young rabbits. After another 2 days the patient was discharged from hospital under outpatient supervision. When the control ultrasound, performed 6 weeks after transplanting patterns. Conclusion: pathological changes it is not revealed.

Subsequent 16-month observation of signs of relapse fibromatosis uterus is not found.

Example 2. Patient B. C., 39 years. Suffer from chronic prostatitis for 8 years. Over the last 4 years he began to notice a significant weakening of sexual function. Repeated courses of traditional treatment at the urologist and sexologist no positive effect. The patient was performed by intramuscular transplantation of material containing cells of the prostate obtained from adult rabbits. Already 2 weeks after transplantation, the patient began to notice a significant improvement in General condition, increase efficiency. Over the next 2 months in a patient with increased sexual activity: frequency of full sexual intercourse increased from 2.1 per month to 2.5 per week. Such a significant improvement in sexual function were observed for at least 8 months. after transplantation.

1. The method of obtaining material containing cells of the prostate gland, which consists in using as source material the prostate gland of a mammal, with the extracted prostate washed, the (growth) environment and serum mammals, followed by incubation in an incubator at a constant temperature, followed by exposure at a lower temperature, characterized in that the extraction of prostate cells from slices of the prostate gland provide the mode of incubation at a constant temperature in the range 35 - 40oC for from two days to six weeks once the temperature drops to 4 - 25oC and holding at that temperature, as well as one-time or periodic reduction in serum concentration of mammals, and then collect the cells of the prostate gland with the culture of the mattress and get the material for transplantation.

2. The method according to p. 1, characterized in that the washing of the prostate using cold Hanks solution with the addition of antibiotics.

3. The method according to p. 1, characterized in that before chopping prostate remove the capsule of the gland and the layer of connective tissue with blood vessels.

4. Material containing cells of the prostate obtained from the prostate gland of a mammal, characterized in that it is obtained by the method according to p. 1.

5. A method of treating fibromatosis uterus is element using the material, containing cells of the prostate, p. 4 and carry out its transplantation.

6. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted intramuscularly.

7. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted into the straight muscle of the abdomen.

8. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted into the liver parenchyma.

9. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted via the portal vein or its branches.

10. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted into the pulp of the spleen.

11. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted in the peritoneal cavity.

12. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted into the greater omentum.

13. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted in the splenic artery.

15. The method according to p. 5, characterized in that the material containing cells of the prostate gland, transplanted subcutaneously.

16. The method of treatment of chronic prostatitis, including the introduction into the body of biologically active elements, characterized in that as a biologically active element using a material containing cells of the prostate, p. 4 and carry out its transplantation.

17. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted intramuscularly.

18. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted into the straight muscle of the abdomen.

19. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted into the liver parenchyma.

20. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted via the portal vein or its branches.

21. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted into the pulp of the spleen.

22. The method according to p. 16, characterized in that the material containing the cells prestate the holding cells of the prostate gland, transplanted into the greater omentum.

24. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted in the splenic artery.

25. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted in a specially created muscle pocket.

26. The method according to p. 16, wherein the material containing cells of the prostate gland, transplanted subcutaneously.

27. A method of treatment of disorders of male sexual function, including the introduction of biologically active elements, characterized in that as a biologically active element using a material containing cells of the prostate, p. 4 and carry out its transplantation.

28. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted intramuscularly.

29. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted into the straight muscle of the abdomen.

30. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted into the liver parenchyma.

31. The method according to p. 27, otlichie branches.

32. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted into the pulp of the spleen.

33. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted in the peritoneal cavity.

34. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted into the greater omentum.

35. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted in the splenic artery.

36. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted in a specially created muscle pocket.

37. The method according to p. 27, characterized in that the material containing cells of the prostate gland, transplanted subcutaneously.

 

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