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Use of human amniotic epithelial cells in treatment of autoimmune disease

A technology of amniotic membrane epithelial cells and autoimmunity, applied in allergic diseases, cell culture active agents, epidermal cells/skin cells, etc., can solve problems such as side effects, poor therapeutic effect, and obvious autoimmune diseases

Pending Publication Date: 2019-08-06
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to solve the technical problem that the current treatment methods for autoimmune diseases have obvious side effects or poor therapeutic effects, the present invention provides a therapeutic drug or method for treating autoimmune diseases using amnion epithelial cells

Method used

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  • Use of human amniotic epithelial cells in treatment of autoimmune disease
  • Use of human amniotic epithelial cells in treatment of autoimmune disease
  • Use of human amniotic epithelial cells in treatment of autoimmune disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0076] Example 1-1 Construction of an experimental autoimmune thyroiditis model

[0077] 1. Purchase and feeding of experimental animals

[0078] 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.

[0079] 2. Experimental drugs

[0080] (1) Complete Freund's adjuvant (complete Freund's adjuvant, CFA) American Sigma Company

[0081] (2) Incomplete Freund's adjuvant (IFA) Sigma, USA

[0082] (3) Porcine thyroglobulin (pTg) Sigma, USA

[0083] 3. Grouping of experimental animals

[0084] Grouped according to different treatment times (Table 2-1), 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) and 3 rats in the simp...

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; 6ml L-Glutamine; 6ml Sodium Pyruvate; 6ml MEMNEAA; 600μl 2-ME; 2000× EGF and 100× P / S before use ;

[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 mother (HIV, syphilis, hepatitis A, hepatitis B, hepatitis C and other serological reactions were all negative) after caesarean 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 sterile PBS solution added with double antibody (P / S) to 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 bath at ...

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Abstract

The invention relates to a use of human amniotic epithelial cells (hAECs) in treatment of autoimmune disease. The invention discloses a method for treating and / or improving the autoimmune disease by using effective dose of the amniotic epithelial cells or cell preparations containing the amniotic epithelial cells alone or in combination with other drugs. The autoimmune disease includes hashimoto thyroiditis, uveitis, lupus erythematosus and the like. The amniotic epithelial cells can be given to patients by local injection or intravenous injection, the dosage range given at each time is about10<3>-10<9> cells, the shortcomings of conventional methods are made up to a certain extent, a good therapeutic 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 genetics 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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IPC IPC(8): A61K35/50A61P37/02A61P5/14A61P27/02C12N5/073
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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