Therapeutic application of human amniotic epithelial cells in autoimmune diseases

A technology of amniotic membrane epithelial cells and autoimmunity, applied in allergic diseases, artificial cell constructs, cell culture active agents, etc., can solve the problems of side effects, poor treatment effect, etc., to avoid immune rejection, reduce sources, Effects from a wide range of sources

Pending Publication Date: 2022-05-13
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In summary, in view of the complex and diverse pathogenesis of autoimmune diseases such as thyroiditis, uveitis, and lupus erythematosus, the specific pathogenesis is still not fully understood, and the existing treatment methods such as hormones and immunosuppressants have obvious side effects or treatment Therefore, it is of great significance to find a safer, more effective and economical treatment method to improve the quality of life of patients and reduce the mortality or disability rate.

Method used

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  • Therapeutic application of human amniotic epithelial cells in autoimmune diseases
  • Therapeutic application of human amniotic epithelial cells in autoimmune diseases
  • Therapeutic application of human amniotic epithelial cells in autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0075] Example 1-1 Construction of an experimental autoimmune thyroiditis model

[0076] 1. Purchase and feeding of experimental animals

[0077] CBA / J mice, SPF grade, 30-40g, female, were provided by Shanghai Southern Model Animal Center. Raise in separate cages. The same group of mice were placed in the same cage. Six mice per cage were raised in the Experimental Animal Center of Zhejiang University. Food and water were freely available.

[0078] 2. Experimental drugs

[0079] (1) Complete Freund's adjuvant (complete Freund's adjuvant, CFA) American Sigma Company

[0080] (2) Incomplete Freund's adjuvant (IFA) Sigma, USA

[0081] (3) Porcine thyroglobulin (pTg) Sigma, USA

[0082] 3. Grouping of experimental animals

[0083] Grouped according to different treatment times (Table 1), each group was repeated at least three times, and each experiment consisted of 9 rats in each group, including 3 rats in the model treatment group (EAT+hAECs group), 3 rats in the simple mo...

Embodiment 1-2

[0090] The preparation of embodiment 1-2 human amnion cell experiment solution

[0091] 1. Preparation of amnion epithelial cell culture medium: add 56ml KSR to 500ml DMEM / F12; 6ml L-Glutamine; 6ml Sodium Pyruvate; 6ml MEMNEAA; 600μl 2-ME; 2000× EGF and 100× P / S Add to;

[0092] 2. Preparation of 2000×EGF: Add 1ml of sterile ddH2O to the EGF packaging tube, let it stand for 5-10min to dissolve, then add 4ml of diluent (5% Trehalose PBS), mix well and then distribute to 1.5ml EP tube 100μl per tube;

[0093] 3. Preparation of digestion termination solution: DMEM / F12+10% FBS;

[0094] 4. Preparation of cryopreservation solution: 40% FBS + 50% culture medium + 10% DMSO.

Embodiment 1-3

[0095] Example 1-3 Isolation of human amniotic epithelial cells

[0096] 1. The source of human amniotic membrane

[0097] After the mother’s authorization and consent, the placental tissue of the healthy parturient (HIV, syphilis, hepatitis A, hepatitis B, hepatitis C and other serological reactions were all negative) after cesarean section was taken, the placenta was cut with a cross knife, and the whole amniotic membrane was obtained by mechanical separation.

[0098] 2. Isolation of hAECs

[0099] Wash the amniotic membrane three times with the sterile PBS solution that has added double antibody (P / S), wash away blood and other impurities, and transfer the amniotic membrane to a 50ml centrifuge tube.

[0100] Add 10ml of 0.25% trypsin (bathed at 37°C in advance) to digest for 30s, invert 20 times, and transfer the amnion into another 50ml centrifuge tube.

[0101] Add 15ml of 0.25% trypsin (bathed at 37°C in advance) to the centrifuge tube, and after digestion in a water...

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Abstract

The present invention relates to the use of human amniotic epithelial cells (hAECs) in the treatment of autoimmune diseases. The invention discloses a method for treating and / or improving autoimmune diseases by using an effective dose of amniotic epithelial cells or a cell preparation containing the amniotic epithelial cells independently or in combination with other medicines. The autoimmune diseases comprise hashimoto thyroiditis, uveitis, lupus erythematosus and the like. The amniotic epithelial cells can be given to a patient by adopting methods such as local injection or intravenous injection, and the dosage range of each time is about 103-109 cells, so that the defects of the existing method are overcome to a certain extent, a good treatment effect is generated, and a new clinical treatment scheme is provided for current autoimmune diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the therapeutic use of human amniotic epithelial cells in autoimmune diseases. Background technique [0002] Immune disease refers to the disease caused by the imbalance of immune regulation and affecting the immune response of the body. A generalized immune disease also includes structural or functional abnormalities of the immune system caused by congenital or acquired causes. Autoimmune diseases refer to diseases caused by the body's immune response to self-antigens, resulting in damage to its own tissues. Its main feature is that the etiology is unknown, related to heredity and various environmental factors, etc., and the course of the disease is long, with frequent attacks, causing great pain to the lives of patients. Common autoimmune diseases include Hashimoto's thyroiditis, uveitis, and lupus erythematosus. [0003] Among them, Hashimoto's thyroiditis is the mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/50A61P37/02A61P27/02A61P5/00C12N5/071C12N5/073
CPCA61K35/50A61P37/02A61P27/02C12N5/0605C12N2509/00A61P5/00C12N5/0625C12N2509/10C12N2501/115C12N2501/11
Inventor 余路阳张传宇谭冰李金英袁惟芯郭礼和邵小燕刘佳
Owner ZHEJIANG UNIV
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