Menstrual blood-derived endometrial stem cell preparation and preparation method and application thereof

A technology of stem cell preparation and endometrium, applied in the field of regenerative medicine, can solve the problems of slow proliferation, limited usability, strong invasiveness, etc., and achieve the effect of reducing patient pain, good repair effect, and excellent effect

Pending Publication Date: 2020-01-17
NANTONG UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, its availability is limited by its invasiveness, low extraction yield, and slow proliferation.

Method used

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  • Menstrual blood-derived endometrial stem cell preparation and preparation method and application thereof
  • Menstrual blood-derived endometrial stem cell preparation and preparation method and application thereof
  • Menstrual blood-derived endometrial stem cell preparation and preparation method and application thereof

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preparation example Construction

[0031] The preparation method of menstrual blood-derived endometrial stem cell preparation comprises:

[0032] (1) Preparation of platelet-rich plasma (PRP), the specific steps are as follows:

[0033] (1-1) Draw some peripheral blood into a sterile test tube containing anticoagulant;

[0034] (1-2) Perform the first centrifugation at 130g for 10 min. After centrifugation, the blood is divided into three layers: the upper layer is platelet-poor plasma, the main component is fibrinogen, etc.; the middle layer is highly concentrated platelets, and the lower layer is red blood cells; Use a pipette to absorb part of the red blood cells in the upper layer, the middle layer and the adjacent middle layer, and transfer them into another sterile test tube;

[0035] (1-3) Carry out the second centrifugation at 250g for 10 minutes. After centrifugation, the plasma is divided into three layers: the lower layer is a little red blood cells, the upper layer is platelet-poor plasma, and the ...

Embodiment 1

[0043] Preparation of menstrual blood-derived endometrial stem cells

[0044] The first step, primary isolation and culture of menstrual blood-derived endometrial stem cells: the patient uses a menstrual cup to collect about 5ml of menstrual blood on the second day after menstrual cramps, and transfers the cells to a culture medium containing 0.2ml amphotericin B, 0.2 ml penicillin streptomycin, 0.1ml EDTA-Na2 in PBS solution, store at 4°C, dilute with sterile PBS equal volume, and mix well. Use 1.077g / ml Ficoll sorting, density gradient centrifugation (1800rpm / min, 25min) to separate menstrual blood mononuclear cells, absorb the middle buffy coat, add 2 times the volume of PBS solvent, 1500rpm / min, 10min centrifugal wash twice, Remove the supernatant, add an appropriate amount of complete medium (DMEM-F12 culture solution containing 10% autologous serum, 100 U / mL penicillin, and 100 μg / mL streptomycin) by pipetting, resuspend, inoculate into different culture dishes at 37°C, ...

Embodiment 2

[0048] Preparation of PRP

[0049] The first step is to draw some peripheral blood into a sterile test tube containing anticoagulant;

[0050] The second step is to perform the first centrifugation at 130g for 10 minutes. After centrifugation, the blood is divided into three layers: the upper layer is platelet-poor plasma, the main component is fibrinogen, etc.; the middle layer is highly concentrated platelets, and the lower layer is red blood cells. Use a pipette to draw some red blood cells from the upper layer, the middle layer and the adjacent middle layer, and transfer them into another sterile test tube.

[0051] The third step is to perform the second centrifugation at 250g for 10 minutes. After centrifugation, the plasma is divided into three layers: the lower layer is a little red blood cells, the upper layer is platelet-poor plasma, and the layer between the two layers is platelet-rich plasma.

[0052] Step 4: Use a straw to discard most of the supernatant, and tak...

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Abstract

The invention provides a menstrual blood-derived endometrial stem cell preparation, which comprises platelet-rich plasma and menstrual blood-derived endometrial stem cells. A preparation method of themenstrual blood-derived endometrial stem cell preparation comprises (1) preparing platelet-rich plasma; (2) preparing menstrual blood-derived endometrial stem cells; and (3) preparing the menstrual blood-derived endometrial stem cell preparation. The menstrual blood-derived endometrial mesenchymal stem cells are prepared and are matched with the platelet-rich plasma, so that mutual promotion canbe effectively achieved; the menstrual blood-derived endometrial stem cell preparation is suitable for preparing skin injury treatment medicaments, allows a wound to heal faster and better, can repairinjured skin better, can ease the pain of patients, and has excellent effects.

Description

technical field [0001] The invention belongs to the technical field of regenerative medicine, and relates to technologies such as in vitro separation and expansion of menstrual blood-derived endometrial stem cells and preparation of biological preparations, and in particular to a preparation method and application of menstrual blood-derived endometrial stem cell preparations. Background technique [0002] The skin accounts for about 70% of the surface area of ​​the human body and is the largest organ of the human body. It has the functions of protecting the body, perspiration, sensing temperature changes, and participating in the body's immune response. Common skin injuries include wounds, scalds, burns, etc. Among them, refractory skin injuries and scars caused by injuries will reduce the physiological functions of related tissues or organs in the body, and severe scars can even lead to deformities, which bring great harm to patients. The physical and mental torture caused ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K35/16A61P17/02
CPCA61K35/28A61K35/16A61P17/02A61K2300/00
Inventor 董传明王庆华吴勤峰
Owner NANTONG UNIVERSITY
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