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Pharmaceutical application of amniotic epithelial cell conditioned medium

A technology of amniotic membrane epithelial cells and conditioned medium, applied in the field of stem cells, can solve the problems of immune rejection, storage and transportation inconvenience, and achieve the effect of high product stability and low immunogenicity

Active Publication Date: 2021-04-30
上海优祺生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the direct use of stem cell therapy has the disadvantages of inconvenient storage and transportation as well as immune rejection

Method used

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  • Pharmaceutical application of amniotic epithelial cell conditioned medium
  • Pharmaceutical application of amniotic epithelial cell conditioned medium
  • Pharmaceutical application of amniotic epithelial cell conditioned medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Preparation of amniotic membrane epithelial cell conditioned medium eye drops

[0034] 1) Isolation and culture of amniotic membrane epithelial cells: Isolate the amniotic membrane from human placenta tissue, wash it with sterile saline, and obtain primary amniotic membrane epithelial cells by shear separation and trypsin digestion. Culture amnion epithelial cells in medium, when the growth of cell colonies is found, the adherent cells are fused to 70-80%, digested with 0.25% trypsin, and subcultured; the morphological characteristics of amnion epithelial cells are as follows figure 1 shown.

[0035] 2) Collection: continue to culture amnion epithelial cells to the third generation in a medium suitable for culturing amnion epithelial cells, discard the medium when the cell confluence reaches about 90%, wash the cells 3 times with sterile PBS, and replace with The serum-free medium was further cultured for 24 hours, the cells and cell debris were removed b...

Embodiment 2

[0036] Example 2: Therapeutic Effect of Amniotic Epithelial Cell Conditioned Medium Eye Drops on Allergic Conjunctivitis

[0037] Experimental allergic conjunctivitis mouse model is a mouse model of experimental allergic conjunctivitis induced by ragweed pollen. The establishment method is as follows: 6-week-old Balb / c mice, male or female, were inoculated with 50 μg of SRW pollen (Greer Lab) into 5 ml of ImjectAlum (Thermo Scientific) by footpad injection on day 0, and the normal control group was injected with phosphoric acid Saline buffer solution (PBS). The sensitization procedure was repeated on day 5 to enhance the allergic response. From day 10 to day 14, 1.5 mg of SRW pollen (pH 7.2) suspended in 10 μl of phosphate buffered saline (PBS) was topically applied to both eyes of the mice, and the normal control group was administered phosphate buffered saline (PBS) . The method of eye drops is amniotic epithelial cell conditioned medium eye drops (10 μL) topically applie...

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Abstract

The invention provides a pharmaceutical application of amniotic epithelial cell conditioned medium eye drops, an application of the amniotic epithelial cell conditioned medium in preparation of the eye drops and an application of the amniotic epithelial cell conditioned medium in preparation of drugs for allergic conjunctivitis. The invention also discloses a preparation method of the amniotic epithelial cell conditioned medium eye drops. According to the pharmaceutical application of the amniotic epithelial cell conditioned medium eye drops, it is found and proved for the first time that the conditioned medium derived from human amniotic epithelial cells can inhibit allergic conjunctivitis. When the amniotic epithelial cell conditioned medium eye drops are used for treating conjunctivitis allergic conjunctivitis, the immunogenicity is low, direct use of cells is avoided, and the amniotic epithelial cell conditioned medium eye drops are safe, effective and high in product stability.

Description

technical field [0001] The invention relates to the application of a conditional culture medium of amniotic membrane epithelial cells in the preparation of eye drops. The invention also relates to the application of amnion epithelial cell conditioned medium in the preparation of medicines for allergic conjunctivitis, which belongs to the field of stem cell technology. Background technique [0002] Allergic diseases are a series of diseases caused by hypersensitivity triggered by the immune system to typical harmless substances in the environment, affecting 20-30% of the population, and have become one of the world's major health problems. The proportion used for the treatment of such diseases is increasing year by year. Typical allergic diseases include hay fever, food allergy, atopic dermatitis, allergic asthma and anaphylaxis. Allergic conjunctivitis (ACD) is such a type of allergic disease that occurs in the conjunctiva of the ocular surface. Clinically, the disease is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/50A61K9/08C12N5/073A61P27/02A61P29/00A61P37/08
CPCA61K35/50A61K9/08A61K9/0048C12N5/0605A61P27/02A61P29/00A61P37/08C12N2500/84C12N2500/90
Inventor 徐国彤徐辉明吴彬欣高芙蓉
Owner 上海优祺生物医药科技有限公司
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