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Application of bronchial basal layer cells in preparation of medicine for treating COPD (chronic obstructive pulmonary disease)

A bronchial and basal layer technology, applied in the field of regenerative medicine, can solve the problems of lack of bronchial basal layer cells, organs or tissues, increasing the risk of opportunistic infections, etc., to achieve large proliferation, low endotoxin content, low chain The effect of mycin residues

Pending Publication Date: 2022-03-18
REGEND THERAPEUTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 1. Seriously insufficient sources of organs or tissues;
[0009] 2. Immunological rejection is obvious. Organ or tissue transplantation is mainly allogeneic transplantation, which often leads to various degrees of immune rejection. If immunosuppressant treatment is used, the risk of various opportunistic infections will increase;
[0010] 3. The operation time is long, the technical requirements are high, and the process is complicated, etc.
[0021] Currently, no clinical-grade bronchial basal cells are available in the prior art

Method used

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  • Application of bronchial basal layer cells in preparation of medicine for treating COPD (chronic obstructive pulmonary disease)
  • Application of bronchial basal layer cells in preparation of medicine for treating COPD (chronic obstructive pulmonary disease)
  • Application of bronchial basal layer cells in preparation of medicine for treating COPD (chronic obstructive pulmonary disease)

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0079] The isolated active bronchi were collected for brush examination for later use. In this embodiment, the tissues located in two different parts of the bronchi were selected. The reason why two or more parts are selected is to ensure the success rate of separation.

[0080] Take the tissue digestion solution and stop solution for later use; 99v% of the tissue digestion solution is DMEM / F12, the rest is 1-20ng / mL DNase, 0.1-4mg / mL protease XIV and 10-200ng / mL trypsin; stop 90v% of the liquid is DMEM, and 10v% is FBS.

[0081] The above-mentioned brushed tissue is digested with the tissue digestion solution, and the digested tissue is terminated with the stop solution, and then the cells are collected.

[0082] Take the medium for later use, in which 225mL DMEM, 225mL F12, 20-70mL FBS, 0.2-2mM L-glutamine (L-glu...

Embodiment 2

[0093] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0094] The isolated active bronchi were collected for brush examination for later use. In this embodiment, the tissues located in two different parts of the bronchi were selected. The reason why two or more parts are selected is to ensure the success rate of separation.

[0095] Take the tissue digestion solution and stop solution for later use; 99v% of the tissue digestion solution is DMEM / F12, the rest is 1-20ng / mL DNase, 0.1-4mg / mL protease XIV and 10-200ng / mL trypsin; stop 90v% of the liquid is DMEM, and 10v% is FBS.

[0096] The above-mentioned brushed tissue is digested with the tissue digestion solution, and the digested tissue is terminated with the stop solution, and then the cells are collected. The cells after enzymatic hydrolysis were tested for bacteria and mycoplasma, and some of them were frozen.

...

Embodiment 3

[0110] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0111] The isolated active bronchi were collected for brush examination for later use. In this embodiment, the tissues located in two different parts of the bronchi were selected. The reason why two or more parts are selected is to ensure the success rate of separation.

[0112] Take the tissue digestion solution and stop solution for later use; 99v% of the tissue digestion solution is DMEM / F12, the rest is 1-20ng / mL DNase, 0.1-4mg / mL protease XIV and 10-200ng / mL trypsin; stop 90v% of the liquid is DMEM, and 10v% is FBS.

[0113] The above-mentioned brushed tissue is digested with the tissue digestion solution, and the digested tissue is terminated with the stop solution, and then the cells are collected. The cells after enzymatic hydrolysis were tested for bacteria and mycoplasma, and some of them were frozen.

...

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Abstract

The invention discloses a preparation method of clinical-grade autologous bronchial basal layer cells and application of a corresponding clinical-grade autologous bronchial basal layer cell reinfusion preparation in treatment of chronic obstructive pulmonary disease (COPD). The preparation method comprises the following steps: taking an in-vitro active bronchus brush inspection tissue, carrying out digestion treatment, and collecting cells after stopping digestion; and carrying out planking culture on the digested cells by using a culture plate paved with trophoblast cells, collecting the cells, carrying out multiplication culture on the cells by using the culture plate paved with trophoblast cells, and carrying out subculture when the cells grow to 50-90% of the surface area of the culture plate. Clinical-grade cells suitable for lung injury repair are determined for the first time, industrial preparation of the cells is achieved through the preparation process, bronchial basal layer cells meeting the clinical cell treatment number and quality can be obtained in a short time, the bronchial basal layer cells prepared through the method can be stably differentiated after entering the focus, and the treatment effect is good. The repair of the lung injury of the COPD patient is realized, and the lung function indexes such as FEV1, FEV1 / FVC, DLCO, MMEF and MVV are improved.

Description

technical field [0001] The invention relates to the technical field of regenerative medicine, in particular to a preparation process of clinical-grade autologous bronchial basal layer cells and its application in treating chronic obstructive pulmonary disease (COPD). Background technique [0002] The lung is a complex organ composed of epithelial cells and other types of cells, which plays an important role in respiration. In the process of performing breathing tasks, especially when exposed to some external or internal harmful substances, such as air pollutants, bacterial viruses, or toxic substances in the blood, a large number of cells in the lungs will die and induce an inflammatory response, thereby Causes massive lung tissue damage. Such massive damage often triggers various lung diseases, such as chronic obstructive pulmonary disease (COPD), bronchiectasis (BE), and pulmonary fibrosis, among others. [0003] Chronic obstructive pulmonary disease (referred to as "chr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/42A61P11/00
CPCC12N5/0688A61K35/42A61P11/08C12N2500/32C12N2501/11C12N2501/33C12N2500/40C12N2501/39C12N2533/90C12N2509/00A61P11/00A61L27/3804A61L27/3839
Inventor 张婷
Owner REGEND THERAPEUTICS CO LTD
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