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Endometrial stem cell preparation for treating erectile disorder and preparation method thereof

A technology of erectile dysfunction and stem cell preparation, applied in the field of biomedicine, can solve the problems of ineffective treatment of erectile dysfunction and psychological burden, and achieve the effect of improving vascular microcirculation, effective treatment, and excellent practical effect

Active Publication Date: 2020-12-08
伯仕利生物科技发展(盐城)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, some V-type phosphodiesterase (phospho-diesterase typev) inhibitor drugs and hormone drugs are widely used in the treatment of impotence. The treatment effect of erectile dysfunction caused by the disease is not obvious, but it will cause psychological burden due to its serious side effects

Method used

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  • Endometrial stem cell preparation for treating erectile disorder and preparation method thereof
  • Endometrial stem cell preparation for treating erectile disorder and preparation method thereof
  • Endometrial stem cell preparation for treating erectile disorder and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0049] Example 1: Culture of endometrial stem cells:

[0050] Implant the menstrual cup on the second day of the menstrual period, continuously collect the menstrual blood for 12 hours, transfer the obtained menstrual blood into an anticoagulant, add penicillin / streptomycin and amphotericin B, and then add normal saline to make a suspension;

[0051] Take the lymphocyte separation medium, spread the suspension on the upper layer of the lymphocyte separation medium, the ratio of the lymphocyte separation medium to the suspension is 1:3, centrifuge at 800g for 20min, and obtain a white film between the lymphocyte separation medium and the suspension layer;

[0052] Take the buffy coat, add normal saline, centrifuge at 2000rpm for 5min, discard the supernatant, then add normal saline, centrifuge at 2000rpm for 5min, discard the supernatant, suspend the cell pellet in the culture medium, and culture in a 37°C incubator; The culture medium was changed every 2 days, cultured for 16...

example 2

[0054] Get the endometrial stem cells prepared in Example 1, and carry out the following identification detection experiments:

[0055] ① Take the cultured endometrial stem cells, centrifuge and suspend them in 1×PBS, count them on a cell counter, and incubate the cells with CD45, CD34, CD90, CD105, and 7-AAD. All antibodies are directly labeled; after incubation, collect the cells , centrifuged and resuspended in 1×PBS, and detected by flow cytometry;

[0056] The channel setting conditions of the flow cytometer are: CD90FITC: 455V; CD45FITC: 455V; CD105PE: 730V; CD34PE: 730V; 7AAD: 876V. The emission wavelengths are: 488nm for FITC; 570nm for PE; 647nm for 7AAD. Tested separately by dosage group.

[0057] ②The cultured endometrial stem cells were taken, and the endotoxin was tested using Zhanjiang Anders Limulus reagent;

[0058] ③Take the cultured endometrial stem cells for fungal and bacterial detection;

[0059] The culture method was used for detection, and the cell ...

example 3

[0065] Example 3: Cryopreservation of endometrial stem cells:

[0066] Take the qualified endometrial stem cells prepared in Example 1, remove the culture medium, and wash the cells twice with PBS; add mild enzymes to digest the cells, shake the culture flask, put it back in the incubator for 1-2min, and observe the cells with an inverted microscope to ensure that all Cells are all detached and floating, tap the side of the flask to release residual adherent cells;

[0067] Then add PBS to dilute with mild enzyme to stop the digestion; take out 100μL for counting; centrifuge the remaining cells at 1000rpm for 5min, and put them in the cryopreservation solution at 1×10 6 Cells / mL density to resuspend the cells, pipette the cell suspension and transfer it to a cryopreservation tube, place it at -80°C overnight, and transfer it to a liquid nitrogen tank for storage.

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Abstract

The invention discloses an endometrial stem cell preparation for treating erectile disorder and a preparation method thereof. Stem cells are pluripotent cells with self-replication capacity, and the stem cells can be differentiated into multiple functional cells under certain conditions. Through stem cell transplantation, damaged cells are replaced by direct differentiation or a large number of cell factors are secreted in a paracrine mode, so that the treatment process is simplified, which is different from the mode that a medicine only aims at specific molecules or signal channels, and therefore, endometrial stem cells which are tissue repair cells with vascular regeneration and repair activity are cultured in vitro. By adopting a micro-injection planting technology, active cells are planted in a male cavernous body in a targeted manner, so that on one hand, the generation of new small vessels can be promoted, and on the other hand, damaged and blocked arterioles can be repaired, themicrocirculation of blood vessels is improved, and the blood supply speed and blood supply quantity of dorsal arteries to the cavernous body are increased; and a safe and effective treatment means isprovided for treating erectile disorder.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an endometrial stem cell preparation and a preparation method for treating erectile dysfunction. Background technique [0002] Erectile dysfunction (Erectile disorder, ED), commonly known as impotence (Impotence), is a typical male sexual dysfunction, usually refers to adult men in the case of sexual stimulation and desire, the penis (for at least 6 months) can not Erection or erection is not firm, and the erection time is short, so that it cannot be inserted into the vagina to complete the sexual intercourse process. [0003] The etiology of erectile dysfunction is more complex, involving many factors such as nerves, endocrine, vascular system, hormone level, psychological state, etc., about 60% of which are mainly caused by mental and psychological factors, and about 30% have organic diseases. Erectile dysfunction caused by organic disorders or diseases may be treated with...

Claims

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Application Information

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IPC IPC(8): A61K35/48A61K47/02A61K47/42A61P15/10C12N5/071A01N1/02
CPCA01N1/02A61K9/0019A61K9/0034A61K35/48A61K47/02A61K47/42A61P15/10C12N5/0682C12N2501/115C12N2509/00C12N2509/10
Inventor 吴金芸叶华衍
Owner 伯仕利生物科技发展(盐城)有限公司
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