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Stem cell exosome preparation for preventing and treating male sexual dysfunction

A technology of stem cells and sexual function, applied in the direction of medical preparations containing active ingredients, diseases, organic active ingredients, etc., can solve the problems of no literature reports and achieve the effect of enhanced effect

Inactive Publication Date: 2021-07-27
奥启(深圳)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no literature report on the combination of cryptotanshinone and mesenchymal stem cells in the treatment of male non-obstructive azoospermia

Method used

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  • Stem cell exosome preparation for preventing and treating male sexual dysfunction
  • Stem cell exosome preparation for preventing and treating male sexual dysfunction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of human bone marrow mesenchymal stem cell exosomes (BMSCs-exo)

[0019] Wash the BMSCs cultured with serum-containing medium for 3 times with PBS; add serum-free medium, collect the culture supernatant after 48 hours; centrifuge at 100g at 4°C for 10min, and take the supernatant; centrifuge at 2000g, 4°C for 20min, and take the supernatant; Filter with a 0.22 μm membrane; centrifuge at 100,000 g at 4°C for 70 min to remove the supernatant; add an appropriate amount of PBS to blow and wash, then centrifuge at 100,000 g at 4°C for 70 min to remove the supernatant; resuspend in an appropriate amount of PBS, and store at -80°C.

[0020] The circular exosome particles wrapped by lipid bilayer membrane were observed by transmission electron microscopy; the positive rate of expression of specific markers CD81 and CD9 protein molecules on the surface of exosomes was detected by flow cytometry; (NTA) analysis results show that the ratio of particle number ...

Embodiment 2

[0021] Example 2 Human bone marrow mesenchymal stem cell exosomes (BMSCs-exo) loaded with cryptotanshinone (CTan)

[0022] After the BMSCs-exo prepared in Example 1 was thawed and stored in a constant temperature bed at 37°C, 50 μg of CTan was mixed with an equal mass of BMSCs-exo, and BMSCs-exo-CTan was prepared by electroporation. The electric shock conditions were voltage 420V, capacitance 150μF, and electroporation was performed in a 4mm electroporation cup. Then, it was filtered with an inverted centrifugal ultrafiltration membrane to remove free compounds that were not transfected into exosomes, and BMSCs-exo-CTan was prepared.

[0023] Establishment of mouse models and grouping

[0024] Get 90 6-7 weeks old male Kunming mice, divide them into experimental group 1-7, model group and control group, wherein experimental group 1-7 and model group were intravenously injected cyclophosphamide solution 60mg / kg / d, control The same amount of normal saline was injected into the...

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Abstract

The invention provides a stem cell exosome preparation for preventing and treating male sexual dysfunction, the active ingredient of the stem cell exosome preparation is obtained by loading cryptotanshinone on exosome derived from mesenchymal stem cells, and the exosome is loaded with cryptotanshinone through an electrotransfection method. In the process of researching the mesenchymal stem cell exosome in the treatment of non-obstructive azoospermia, a researcher finds that the effect of using the mesenchymal stem cell-derived exosome alone was not obvious, but unintentionally finds that the effect is obviously enhanced after cryptotanshinone is loaded into the exosome through the electrotransfection method; in addition, the researcher also finds that different concentrations of exosomes loaded with cryptotanshinone have different effects.

Description

technical field [0001] The invention belongs to the field of stem cell preparations, and in particular relates to a stem cell exosome preparation for preventing and treating male sexual dysfunction. Background technique [0002] Azoospermia is divided into obstructive azoospermia and non-obstructive azoospermia. Among them, non-obstructive azoospermia (NOA) is one of the most common diseases leading to male infertility, and its lesion is located in the testis. The testicles cannot produce sperm, or the testicular spermatogenic ability is low, resulting in no sperm in the semen test. [0003] Exosomes secreted by mesenchymal stem cells (MSCs) are nanoscale cystic vesicles with a double-membrane structure secreted by active cells into the external environment. It is a kind of adult stem cell rich in cell-derived mRNA, miRNA, protein and other components, which has a strong potential to differentiate into various tissue cells. Studies have found that MSCs-exo has anti-inflamm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P15/08A61K31/58
CPCA61K31/58A61K35/28A61P15/08A61K2300/00
Inventor 宋芸娟
Owner 奥启(深圳)生物科技有限公司
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