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Application of mesenchymal stem cells in preparation of products for treating rheumatoid arthritis

A mesenchymal stem cell and rheumatoid technology, which can be applied to the application field of mesenchymal stem cells in the preparation of products for the treatment of rheumatoid arthritis, and can solve the problem that the pathogenesis is not very clear.

Inactive Publication Date: 2020-05-29
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the pathogenesis of RA is not very clear, there is no cure

Method used

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  • Application of mesenchymal stem cells in preparation of products for treating rheumatoid arthritis
  • Application of mesenchymal stem cells in preparation of products for treating rheumatoid arthritis
  • Application of mesenchymal stem cells in preparation of products for treating rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1. Preparation of mesenchymal stem cell preparations from Wharton's umbilical cord

[0060] 1. Obtaining the umbilical cord of the fetus

[0061] Take the isolated umbilical cord of a full-term newborn without congenital disease; the parturient does not have infectious diseases such as hepatitis, syphilis, AIDS, and the parturient and family members have informed consent to the use of the umbilical cord for experimental research.

[0062] 2. Obtaining mesenchymal cells in the Wharton area of ​​the umbilical cord

[0063] 1. Pretreatment of umbilical cord

[0064] In a sterile laboratory bench, the umbilical cord is repeatedly rinsed with normal saline to wash away residual blood. Cut the umbilical cord into 2-3cm sections with sterile surgical instruments, cut the umbilical cord longitudinally, remove the umbilical artery, umbilical vein and amniotic membrane, take the Wharton area and cut it into 0.5-1mm 3 Small pieces around.

[0065] 2. Obtaining primary umbilical cord...

Embodiment 2

[0077] Example 2. Identification of P3 cell preparation

[0078] The test product is the P3 cell preparation prepared in Example 1.

[0079] Use flow cytometry to detect the expression of CD29, CD44, CD90, CD105, CD126, CD31, CD45, CD34 and HLA-DR in the test product. The specific steps are as follows: After the cells (test product) are incubated with the corresponding antibodies respectively , Wash off excess antibodies, resuspend the cells in PBS buffer, and use BD’s LSR Fortessa instrument to detect the positive rate of each indicator.

[0080] The test results of mesenchymal stem cell markers (CD29, CD44, CD90, CD105 and CD126) are shown in figure 1 . For the test results of hematopoietic stem cell markers (CD31, CD45 and CD34), see figure 2 . For the test results of the markers related to transplant rejection (HLA-DR), see image 3 . The results showed that the test product expressed high expression of mesenchymal stem cell markers (both positive rates were above 97%), low e...

Embodiment 3

[0081] Example 3. The therapeutic effect of P3 cell preparation on rheumatoid arthritis

[0082] 1. Model preparation and administration

[0083] Test mice: 6-8 weeks, DBA / 1J mice, adapted to feed for one week (clean animal breeding room, free drinking water). Chicken type II collagen and Freund's adjuvant were used to sensitize mice to induce arthritis model. Divided into the following groups according to whether it is administered or not and the difference in administration:

[0084] Normal control group (5 mice): 0.1 mL of saline was used to subcutaneously inject each mouse's tail root. After 3 weeks, 0.1 mL of normal saline was injected subcutaneously into the base of each mouse's tail. From the day of the second injection, normal saline was injected into the tail vein once a week (100 μL per injection).

[0085] Cell therapy group (8 animals): use 0.1 mL of emulsified antigen adjuvant suspension (mixed with the same volume of 2 mg / mL chicken type II collagen solution and Freun...

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Abstract

The invention discloses application of mesenchymal stem cells in preparation of products for treating rheumatoid arthritis. The invention provides application of the mesenchymal stem cells or cell preparations of the mesenchymal stem cells in preparation of drugs for treatment of rheumatoid arthritis. The preparation method of the cell preparations includes the following steps: adopting a tissue block in a Warton zone of the umbilical cord as a raw material, and preparing the umbilical cord mesenchymal stem cells so as to obtain the cell preparations, wherein a low-serum culture medium is adopted in the preparation process. The cell preparations are suitable for cell therapy of rheumatoid arthritis and other diseases related to rheumatoid arthritis, wherein complications and diseases withsimilar pathogenesis of the diseases related to rheumatoid arthritis include rheumatoid vasculitis, pneumonia, osteoarthritis, rheumatoid heart disease, ankylosing spondylitis, gout, eye diseases, kidney diseases, urinary system infection, Cushing's syndrome, oral ulcers and other diseases. A great application value is provided for treatment of rheumatoid arthritis and related diseases.

Description

Technical field [0001] The invention relates to the application of mesenchymal stem cells in the preparation of products for treating rheumatoid arthritis, and in particular to the application of human umbilical cord Wharton mesenchymal stem cell preparations in the preparation of products for treating rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by joint synovitis and articular cartilage erosion and destruction. The global incidence rate is about 1%. A sample survey in my country The result is about 0.8%; its main pathological manifestations include three aspects: abnormal immune function, synovial hyperplasia, and destruction of articular cartilage and bone tissue. The pathological changes of joints are mainly manifested by inflammatory cell infiltration, synovial hyperplasia, pannus formation, cartilage and synovial damage, etc.; pannus gradually extends from the synovial membrane at the edge of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61P19/02A61P37/02C12N5/0775
CPCA61K35/51A61P19/02A61P37/02C12N5/0668A61K35/28A61K38/16A61P29/00
Inventor 常智杰付艳霞盛剑秋任芳丽王银银韩斌冯亚瑞
Owner TSINGHUA UNIV
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