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Use of human amniotic epithelial cells in treatment of graft-versus-host disease

An amniotic epithelial cell and graft-versus-host technology is applied in the biological field to achieve the effects of reducing target organ lesions, reducing infiltration, and achieving good results.

Active 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

[0009] The clinical application of Allo-HSCT has made great progress due to the optimization of donor selection, improvement of conditioning regimen, optimization of graft-versus-host disease (GVHD) prevention regimen, and improvement of supportive treatment methods. However, acute graft-versus-host disease (aGVHD) is still one of the leading causes of death after transplantation, and the development of new clinical treatments is urgently needed

Method used

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  • Use of human amniotic epithelial cells in treatment of graft-versus-host disease
  • Use of human amniotic epithelial cells in treatment of graft-versus-host disease
  • Use of human amniotic epithelial cells in treatment of graft-versus-host disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Isolation and culture of primary amniotic membrane epithelial cells

[0058] 1. The source of human amniotic membrane

[0059] In order to avoid microbial contamination of the birth canal, fetal placenta was selected by caesarean section. Due to the stimulation of labor signals after term, the amniotic membrane will undergo apoptosis, so it is advisable to use premature fetal placenta (before 38 weeks). 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.

[0060] 2. Isolation of hAECs (aseptic operation is required throughout the process)

[0061] The placenta of infants delivered by caesarean section before 39 weeks was obtained, the amniotic membrane was removed from the i...

Embodiment 2

[0073] Example 2 Separation, cultivation and cryopreservation of human peripheral blood mononuclear cells

[0074] Aseptically draw the donor's peripheral blood into a collection strip containing anticoagulant.

[0075] Add 5ml of Ficoll solution to a 50ml centrifuge tube, draw blood with a sterile Pasteur pipette and gently drop on the Ficoll solution.

[0076] Without centrifugal acceleration, centrifuge at 400g for 30min at room temperature until the blood is clearly stratified.

[0077] Carefully absorb the buffy coat layer in the stratification with a new Pasteur pipette, add it to 10ml of DPBS (containing 1xPS) and mix well. Centrifuge at 300g for 5min at room temperature and discard the liquid.

[0078] Resuspend the pellet in 10ml of DPBS (containing 1xPS), centrifuge at 300g for 5min at room temperature, and discard the liquid.

[0079] Resuspend in complete medium (1640, 10% FBS, PS) for later use, or resuspend in freezing solution and freeze in liquid nitrogen. ...

Embodiment 3

[0080] Example 3 Mouse Model Establishment and Scoring Criteria for aGVHD Mouse

[0081] 1. Establishment of irradiation injury model in NOD / SCID mice

[0082] NOD / SCID mice purchased from the company at the age of 6-8 weeks were transferred to the animal room for 1 week to adapt to the environment. Add gentamicin (32x10^4U / L) and erythromycin (250mg / L) to drinking water.

[0083] One week later, the mice were sublethally irradiated with 2.7 Gray dose of X-rays.

[0084] The next day, 2×10^6 hAECs were injected into each mouse via the tail vein. The control group was injected with corresponding volume of PBS.

[0085] 2. Establishment of acute GVHD model in NCG mice

[0086] NCG mice aged 6-8 weeks were purchased from the company and transferred to the animal room for 1 week to adapt to the environment. Add gentamicin (32x10^4U / L) and erythromycin (250mg / L) to drinking water.

[0087] One week later, each mouse was injected with 10^7 human peripheral blood mononuclear ce...

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Abstract

The invention relates to a use of human amniotic epithelial cells (hAECs) in treatment of graft-versus-host disease (aGVHD). The invention discloses a method for treating and / or improving the graft-versus-host disease by using effective dose of the amniotic epithelial cells or a cell preparation containing the amniotic epithelial cells alone or in combination with other drugs. The amniotic epithelial cells can be given to patients by intravenous injection or intramedullary injection, the dosage range given at each time is about 10<3>-10<9> cells, the infiltration of inflammation cells caused by aGVHD to target organs can be effectively alleviated, the lesion of target organs is also significantly alleviated, the hAECs are found out to have an apoptosis-promoting effect on multiple leukemiacell lines and be used for treating the graft-versus-host disease.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of human amniotic epithelial cells in the treatment of graft-versus-host disease. Background technique [0002] Hematopoietie stem cell transplantation (HSCT) was first introduced into clinical practice by American scientist Professor Thomas in the 1950s. After decades of development, HSCT is used to treat a large number of malignant blood system diseases and metabolic birth defects. The only existing cure for tumors. [0003] In hematopoietic transplantation, graft-versus-leukaemia (GVL) or graft-versus-tumour (GVT) is the main mechanism for its treatment of cancer. In contrast, graft-versus-host disease (GVHD), the main complication after hematopoietic stem cell transplantation, is the leading cause of death after transplantation. According to statistics, about 50% of patients receiving hematopoietic transplantation will develop GVHD symptoms thus seve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/36A61P37/06C12N5/071C12N5/073
CPCA61K35/36A61P37/06C12N5/0605C12N5/0625C12N2501/11C12N2501/115C12N2509/00
Inventor 余路阳张传宇杨棚捷袁惟芯李金英郭礼和邵小燕刘佳
Owner ZHEJIANG UNIV
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