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Application of exosomes derived from menstrual blood stem cells on preparation of medicament for treating intrauterine adhesion

A technology of intrauterine adhesions and stem cells, applied in the field of biomedicine

Active Publication Date: 2019-06-18
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the advantages of exosomes derived from menstrual blood stem cells, exosomes derived from menstrual blood stem cells are expected to be used in the preparation of drugs for the treatment of intrauterine adhesions, but there have been no related reports so far

Method used

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  • Application of exosomes derived from menstrual blood stem cells on preparation of medicament for treating intrauterine adhesion
  • Application of exosomes derived from menstrual blood stem cells on preparation of medicament for treating intrauterine adhesion
  • Application of exosomes derived from menstrual blood stem cells on preparation of medicament for treating intrauterine adhesion

Examples

Experimental program
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Embodiment 1

[0028]Example 1 Preparation of exosomes derived from menstrual blood stem cells.

[0029] 1. Experimental equipment, materials and reagents.

[0030] Ultra clean bench, CO 2 Incubator, inverted microscope, centrifuge, ultracentrifuge, constant temperature water bath, B-ultrasound machine, 25cm 2 Cell culture bottle, centrifuge tube (50 ml, 15 ml), pipette, 5 ml syringe, artificial insemination catheter, blood collection needle, DMEM-F12 culture medium, 0.25% trypsin, phosphate buffer saline (PBS, Ph=7.4, purchase from Biological Industries, Cat. No. 0044818).

[0031] 2. Primary isolation and culture of menstrual blood stem cells.

[0032] (1) Venous blood collection: Come to the hospital on the first day after menstrual cramps, take 30 ml of venous blood from the blood collection needle, let it stand overnight at 4°C, centrifuge at 3000rpm for 10 minutes at room temperature, take serum, inactivate at 56°C for 30 minutes, and store it at -20°C save.

[0033] (2) Menstrual...

Embodiment 2

[0046] Example 2 Study on the mechanism of intrauterine adhesions in rats treated with blood-derived mesenchymal stem cells and exosomes derived from them.

[0047] Obtain endometrium from a small number of patients during hysteroscopic endometrial adhesion release (routine operation, signed informed consent), culture endometrial cells at the adhesion site in vitro, and red fluorescent dye DiI labeled menstrual blood stem cell-derived exosomes , added to the complete medium for 24 hours, found to be internalized by endometrial stromal cells in patients with intrauterine adhesions, such as image 3 shown.

[0048] 1. Establishment of intrauterine adhesion rat model.

[0049] Female SD rats aged 8-10 weeks were selected, and the bilateral endometrial injury model was established by mechanical injury method. The rats were sacrificed by cervical dislocation 9 days after the injury. Uterine tissue was collected, and the histopathological structure was analyzed by H&E staining, an...

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Abstract

The invention belongs to the field of biological medicines, and particularly relates to application of exosomes derived from menstrual blood stem cells on preparation of a medicament for treating intrauterine adhesion. After primary isolation, culture and passage of the menstrual blood stem cells are carried out, 3rd-6th generations of menstrual blood stem cells are selected for exosomes preparation. When the density of the menstrual blood stem cells reaches 80%, serum components are removed by PBS washing, serum-free DMEM-F12 culture medium is added, and the menstrual blood stem cells are cultured in an incubator at the conditions of 37 DEG C, 5% CO2 and saturated humidity for 24 hours. The supernate is collected, the exosomes are prepared by ultracentrifugation under GMP conditions, andsterile PBS is collected and stored at the temperature of 80 DEG C. The cells have the advantages of being high in survival rate, stable in characters and easy to acquire and expand, and having an obvious treatment effect on intrauterine adhesion. The exosomes can even completely simulate the biological function of the cells to achieve the effects of promoting tissue repair and treating diseases.The exosomes are safer than the cells where the exosomes are derived from, do not lose functions after cryopreservation, can be used for allotransplantation and are suitable for mass production, and have important clinical application values and social and economic benefits.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of exosomes derived from menstrual blood stem cells in the preparation of drugs for treating intrauterine adhesions. Background technique [0002] The endometrium plays a pivotal role in maintaining female physiological characteristics and fertility. It has a high regenerative capacity and proliferative activity. Any factor that causes endometrial destruction can cause intrauterine adhesions. With the popularization of frequent uterine operations and hysteroscopic surgery, the incidence and detection rate of intrauterine adhesions (IUA) caused by endometrial injury are increasing year by year, and it has become the second most common cause of secondary infertility in women. Two reasons. IUA is due to the formation of fibrous scars after the endometrial basal layer is damaged, and repair obstacles lead to the partial or complete adhesion of the uterine cavit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/14A61K35/28A61K35/48A61P41/00A61P15/00
Inventor 谭季春李萍萍张斯文常琦圆
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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