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INVENTION
Russian Federation Patent RU2286790
METHOD stress urinary incontinence
Name of the inventor: Kulakov Vladimir (RU); Dry Gennady T. (RU); Balan Vera Efimovna (RU); Smetnik Vera (RU)
The name of the patentee: State Establishment Scientific Center for Obstetrics, Gynecology and Perinatology, Russian Academy of Medical Sciences
Address for correspondence: 117997, Moscow, ul. Acad. Oparin, 4, PG NC Obstetrics, Gynecology and Perinatology, Director
Starting date of the patent: 2005.03.16
The invention relates to the field of medicine, and relates to the treatment of stress urinary incontinence in women through the introduction of fetal stem cells. In rhabdosphincter lacunar injected 40 million stem myoblasts and fibroblasts obtained from the fruit of 8-12 weeks of gestation, at a ratio of 5-4 cells:. 3-2. The method extends the arsenal of treatments for stress urinary incontinence in women, avoids surgery.
DESCRIPTION OF THE INVENTION
The present invention relates to treatment, in particular to methods for treating stress urinary incontinence.
Urogenital disorders in menopause - a set of vaginal and urinary symptoms, the development of which is a complication of atrophic and degenerative processes in estrogen-dependent tissues and structures of the lower third of the urogenital tract: bladder, urethra, vagina, ligaments and muscles of the pelvic floor.
Currently, the only way to treat stress urinary incontinence is surgery, which has more than 200 modifications (H.Hirsh, O.Kezer, F.Ikle "Operative Gynecology" Publishing House GEOTAR Medicine, Moscow, 1999, str.407).
A disadvantage of the method is the high frequency of relapses and complications due to surgery.
We describe an attempt to treat urinary incontinence in 8 patients by transurethral injection into rhabdosphincter and urethra combination of autologous stem cell-derived muscle tissue and connective tissue cells (Shiomo Pas MD "Stem cell offer exciting approach to urinary incontinence treatment" Urology Times 1 July, 2004).
Although the population of pluripotent cells was obtained from the skeletal muscle, but the authors do not lead publication data confirming that the cells used were actually stem, as well as the success of the treatment in the time slot.
The disadvantage of this method is its duration, related to the need of cellular material intake and growing stem cells thereof.
Object of the invention - to simplify the method of treatment of stress urinary incontinence in women by using more affordable allogeneic cell material.
This object is achieved by a method of treatment of urinary incontinence, lies in the fact that the injected allogeneic embryonic or fetal somatic stem cells - myoblasts and fibroblasts at a ratio 5-4: 3-2 in lacunar rhabdosphincter.
Preparation of cell cultures enriched human myoblasts for subsequent transplantation.
Corpse abortive material is brought to the autopsy block in the cold box (+ 4 ° C) no later than 4 hours after the abortion. The fruits were washed with a sterile saline solution with antibiotics. Striated muscle isolated from human fetuses 8-12 weeks gestation, washed with phosphate buffered saline (PBS) pH 7.3 with the addition of antibiotics. Fragments of muscle tissue was mechanically dissected in a sterile petri dish (35 mm) to a size of not more than 1.0 mm × 1,0 mm. Washed twice with PBS, centrifuging at 800 g for 3 minutes. Dissitsiirovali pieces of muscle tissue into individual cells by incubation at 37 ° C with 0.2% collagenase for 15 minutes. Then washed twice PBS.
A population of cells enriched myoblasts were placed into culture flasks at a concentration of 500,000 cells / ml in cell culture medium: F12 and DMEM in a ratio of 1: 1, supplemented with fetal fetal serum, L-glutamine, HEPES, sodium bicarbonate, antibiotics. Cells were cultured at 37 ° C in an atmosphere with 5% CO 2 until the monolayer was removed by passaging the fibroblasts. Changed every 3 days by 50-75%. The suspension enriched myoblasts were cultured by the above scheme until passage 3, are then used for transplantation or frozen by the standard program transfer into liquid nitrogen.
Getting a culture of human fibroblast cells for later transplantation.
Corpse abortive material is brought to the autopsy block in the cold box (+ 4 ° C) no later than 4 hours after the abortion. The fruits were washed with a sterile saline solution with antibiotics. skin tissue isolated from human fetuses 8-12 weeks of gestational age, washed with phosphate buffered saline (PBS) pH = 7.3 with the addition of antibiotics. Skin fragments were subjected to mechanical dissection under sterile Petri dishes (35 mm) to a size of not more than 1.0 mm × 1,0 mm. Washed twice with PBS, centrifuging at 800 g for 3 minutes. Dissitsiirovali pieces of muscle tissue into individual cells by incubation at 37 ° C with 0.2% collagenase for 25 minutes. Then washed twice PBS.
a population of cells enriched fibroblasts were placed into culture flasks at a concentration of 500,000 cells / ml in cell culture: F12 and DMEM in a ratio of 1: 1, supplemented with 10% fetal calf serum, L-glutamine, HEPES, sodium bicarbonate, antibiotics. Cells were cultured at 37 ° C in an atmosphere with 5% CO 2 to obtain a monolayer. Changed every 3 days by 50-75%. The fibroblasts cultured by the above scheme until passage 3, are then used for transplantation or frozen by the standard program transfer into liquid nitrogen.
The following example illustrates the invention.
LK patient, 55 years old, suffering from stress urinary incontinence in 5 years. Cough test is positive, Valsalvy test is positive, a complete urodynamic study - lack of reflex-apparatus of the bladder, reducing the maximum intraurethral pressure by 50%, the treatment of hormone replacement therapy without effect. The patient lacunar in rhabdosphincter introduced 40 million embryonic stem myoblasts and fibroblasts cells at a ratio of 4:. 2.
Observation of the patient after two weeks showed clinical improvement:
1. Holds the urine under a load of (cough, laugh).
2. Sample Valsalvy negative.
Further monitoring of the patient (for 5 months) showed the preservation of the therapeutic effect.
CLAIM
A method for treating urinary stress incontinence in women by administering a rhabdosphincter cellular material, characterized in that the lacunar rhabdosphincter administered stem 40 million myoblasts and fibroblasts derived from fetuses 8-12 weeks gestation cells at a ratio of 5.4:. 3-2.
print version
Publication date 06.01.2007gg
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