Application of amniotic fluid stem cells in preparation of drugs for treating lupus nephritis

A lupus nephritis and stem cell technology, which is applied in the application field of medicine, can solve the problems of donor injury, the decline of the number, proliferation and differentiation ability of mesenchymal stem cells, and achieves the reduction of urine protein level, simple and rapid in vitro culture, and induction of differentiation. powerful effect

Active Publication Date: 2020-10-13
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, animal experiments and clinical applications have confirmed that mesenchymal stem cells derived from bone marrow and umbilical cord can delay the progression of lupus nephritis through immune regulation, but there are two defects that cannot be ignored when using mesenchymal stem cells such as bone marrow and fat to treat diseases : First, the mesenchymal stem cells will cause different degrees of damage to the donor; second, the number, proliferation and differentiation ability of the mesenchymal stem cells will decrease with the age of the donor
However, whether amniotic fluid stem cells can play a therapeutic role in lupus nephritis is still unknown

Method used

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  • Application of amniotic fluid stem cells in preparation of drugs for treating lupus nephritis
  • Application of amniotic fluid stem cells in preparation of drugs for treating lupus nephritis
  • Application of amniotic fluid stem cells in preparation of drugs for treating lupus nephritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Sorting, culturing and identification of amniotic fluid stem cells

[0039] In this experiment, the amniotic fluid stem cells in the amniotic fluid in the second trimester of pregnancy were sorted and purified by means of differential adherence and mechanical separation, and then the amniotic fluid stem cells were identified by flow cytometry to detect surface antibodies and adipogenic osteogenic differentiation, in order to The next step is to use amniotic fluid stem cells for research.

[0040] 1.1 Experimental materials

[0041] 1.1.1 Experimental samples

[0042] Amniotic fluid samples in the second trimester of pregnancy were obtained from the Department of Obstetrics and Gynecology of the General Hospital of the Chinese People’s Liberation Army. On the basis of excluding fetal malformations and maternal diseases, 10 to 15 ml of amniocentesis was performed in the second trimester (referring to the 14th week of pregnancy to the end of 27th week of pregna...

Embodiment 2

[0102] Example 2. Study on the in vivo effectiveness of amniotic fluid stem cells in treating lupus nephritis

[0103] 2.1 Experimental materials

[0104] 2.1.1 Experimental animals

[0105] Thirty 3-4-week-old female lupus nephritis mice (MRL / lpr) mice were purchased from Shanghai Slack Experimental Animal Co., Ltd. Breeding conditions: SPF grade, room temperature 26°C, relative humidity 50%, constant temperature and humidity, light and dark alternately once every 12 hours, mice eat and drink freely in the breeding cage, and the litter is changed every 3 days. This experiment was reviewed by the Hospital Ethics Committee.

[0106] 2.1.2 Experimental reagents

[0107] 1% periodic acid, Schiff's solution, hematoxylin staining solution, ammonia water, xylene, alcohol, umbilical cord mesenchymal stem cells (Tianjin National Stem Cell Center), BCA protein concentration assay kit (Shanghai Biyuntian), GSM-CAA-4000 Kit (see Table 1 below for the content of the kit):

[0108] Ta...

Embodiment 3

[0205] Example 3, the immune regulation and mechanism of amniotic fluid stem cells in lupus nephritis

[0206] 3.1 Experimental materials

[0207] 3.1.1 Experimental specimens

[0208] Each sample was mixed from 2 identically treated MRL / lpr mouse kidneys, and the experiment was divided into 3 groups, with 3 samples in each group, a total of 9 samples. Among them: 3 cases in the CON group, 3 cases in the AF group, and 3 cases in the UC group.

[0209] 3.1.2 Experimental instruments and equipment

[0210] Helios mass spectrometry flow cytometer (FLUIDIGM, USA)

[0211] 3.1.3 Experimental reagents

[0212] 42 mass spectrometry metal isotope-coupled antibodies (see Table 2, used to label different cell subsets (T cells, B cells, NK cells, granulocytes, myeloid cells, etc.) (FLUIDIGM, USA)

[0213] 194Pt (FLUIDIGM, USA), Cisplatin stain (FLUIDIGM, USA), FACS Buffer (FLUIDIGM, USA), Block Solution reagent (FLUIDIGM, USA), Intercalator-Ir stain (FLUIDIGM, USA), Fix and Perm Buf...

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Abstract

The invention provides an application of amniotic fluid stem cells in preparation of drugs for treating lupus nephritis. The amniotic fluid stem cells can down-regulate the expression of pro-inflammatory factors IL-17 and IL-12p40 and up-regulate the expression of an anti-inflammatory factor IL-1ra, and meanwhile, can down-regulate pro-inflammatory M1 macrophages, dendritic cells and eosinophils and up-regulate anti-inflammatory Th2 cells so as to exert the treatment effect. The amniotic fluid cells have the characteristics of being easy to obtain, free of ethical disputes, simple to culture,strong in differentiation capability and low in immunogenicity. A new strategy is provided for treatment of systemic lupus erythematosus and lupus nephritis which is a complication of systemic lupus erythematosus.

Description

technical field [0001] The present invention relates to a new application of amniotic fluid stem cells, specifically, the present invention relates to the application of amniotic fluid stem cells in the preparation of medicines for treating lupus nephritis. Background technique [0002] Systemic Lupus Erythematosus (SLE) is a complex autoimmune disease involving multiple organs. Various factors are closely related to the pathogenesis of SLE, including genetics, drugs, environment, infection, endocrine and mental factors, etc. These factors lead to immune dysfunction, abnormal immune tolerance, and a large number of autoantibodies such as anti-dsDNA antibodies and antinuclear antibodies, which affect various organs and bring great harm to people's health. SLE is more common in female patients (the ratio of male to female is about 1:9), especially in women of childbearing age. Although male patients are less common, disease activity and clinical symptoms are more serious than...

Claims

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Application Information

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IPC IPC(8): A61K35/50A61P37/02A61P13/12
CPCA61K35/50A61P37/02A61P13/12
Inventor 陈香美白雪源张俊铭
Owner GENERAL HOSPITAL OF PLA
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