Clinical-grade autogenous bronchial basal layer cell, transfusion preparation and preparation technology

A preparation process and bronchial technology, applied in the field of regenerative medicine, can solve the problems of lack of bronchial basal layer cells, organs or tissue sources, increased risk of opportunistic infections, etc., to achieve safety and effectiveness, low endotoxin content , the effect of low streptomycin residue

Pending Publication Date: 2020-11-17
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

Method used

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  • Clinical-grade autogenous bronchial basal layer cell, transfusion preparation and preparation technology
  • Clinical-grade autogenous bronchial basal layer cell, transfusion preparation and preparation technology
  • Clinical-grade autogenous bronchial basal layer cell, transfusion preparation and preparation technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0078] 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.

[0079] 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.

[0080] 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.

[0081] Take the medium for later use, in which 225mL DMEM, 225mL F12, 20-70mL FBS, 0.2-2mM L-glutamine (L-glu...

Embodiment 2

[0092] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0093] 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.

[0094] 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.

[0095] 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

[0109] This embodiment provides a preparation process of clinical-grade autologous bronchial basal layer cells, which specifically includes the following steps:

[0110] 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.

[0111] 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.

[0112] 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 clinical-grade autogenous bronchial basal layer cell, a transfusion preparation and a preparation technology in the technical filed of regeneration medicine. The preparationtechnology specifically includes the following steps: taking in-vitro active bronchial brushing tissue to perform digestive treatment, and collecting cells after digestion is finished; performing planking culture on the digested cells by using a culture plate coated with trophoblastic cells, collecting the cells and using the culture plate coated with the trophoblastic cells to perform amplification culture, and performing subculturing when the cells are grown to 50%-90% of the surface area of the culture plate; and performing digestion and collecting adherent cells when the subcultured cellsare grown to 85%-95% of the surface area of the culture dish, and then performing washing. The clinical-grade cells are firstly determined to be suitable for lung injury repair; through the selectionof proper preparation technology, the industrialized preparation of the cells can be realized, and bronchial basal layer cells that meet the quantity and quality of clinical cell treatment can be obtained within a short time; and the bronchial basal layer cells prepared by the method can stably differentiate when entering the focus, so that the obvious repairing of the lungs can be realized.

Description

technical field [0001] The invention relates to the technical field of regenerative medicine, in particular to a clinical-grade autologous bronchial basal layer cell, a reinfusion preparation and a preparation process. 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 "chronic obstructive pulmonary disease", COPD) is a g...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61L27/38
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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