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Mature airway organoids, methods of making and uses thereof

An organoid and airway technology, applied in biochemical equipment and methods, instruments, scientific instruments, etc., can solve problems such as difficult to reproduce human airway epithelial histology, low serine protease activity, and unsupported growth of influenza virus

Pending Publication Date: 2021-03-26
THE UNIVERSITY OF HONG KONG +1
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Problems solved by technology

However, this ability is lost after 2-3 passages (Butler, et al., Am J Respir Crit Care Med194(2):156-168(2016))
In conclusion, these primary tissues and cells constitute few convenient, reproducible models for studying human respiratory pathogens
Although there are various cell lines (such as A549 and MDCK) that are commonly used to propagate influenza virus and study virology, they have difficulty reproducing the histology of human airway epithelium
Furthermore, due to the low activity of serine proteases, most cell lines do not respond unless the medium is supplemented with an exogenous serine protease: trypsin treated with N-tosyl-L-phenylalanine chloromethyl ketone (TPCK). Supports the growth of influenza viruses with a mono-HA cleavage site

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  • Mature airway organoids, methods of making and uses thereof
  • Mature airway organoids, methods of making and uses thereof
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[0022] I. Definition

[0023] As used herein, "basal medium" refers to basal salt nutrients or aqueous solutions of salts and other elements that provide cells with water and certain large quantities of inorganic ions necessary for normal cell metabolism and maintain intracellular and / or extracellular Osmotic balance.

[0024] An ErbB3 / 4 ligand is defined herein as a ligand capable of binding ErbB3 and / or ErbB4.

[0025] As used herein, the term "induced pluripotent stem cell" (iPSC) is a type of pluripotent stem cell artificially derived from a non-pluripotent cell.

[0026] As used herein, "media, culture media" refers to an aqueous-based solution for the growth, survival or storage of cells used in the practice of the present invention. Media can be natural or artificial. The culture medium may comprise basal medium and may be supplemented with nutrients (e.g., salts, amino acids, vitamins, trace elements, antioxidants) to promote desired cellular activities, such as cel...

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Abstract

Provided are methods for generating 2D and 3D differentiated airway organoids, 2D and 3D differentiated airway organoids which are generated by the methods and uses for the 2D and 3D differentiated airway organoids.

Description

technical field [0001] The present invention relates generally to airway organoids, particularly differentiated airway organoids, methods of making and using them, especially for influenza virus research. [0002] Background of the invention [0003] Influenza A virus (IAV) can infect a variety of avian and mammalian species, including humans, and has a remarkable ability to evolve and adapt to new hosts (1). IAV's fragmented RNA genome and low-fidelity RNA polymerase allow antigenic translocation and drift, which drove this evolution. Thus, new viruses from birds and pigs can cross species barriers and infect humans, causing sporadic infections, epidemics, and even pandemics (Klenk, Cell Host Microbe 15(6):653-654(2014); To, et al ., Lancet 381(9881):1916-1925(2013)). Despite tremendous advances in virology and epidemiology, it is still impossible to predict which subtype or strain of IAV will cause the next outbreak. Since 2013, a novel reassortant H7N9 influenza virus f...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N7/00C12Q1/70C12Q1/02
CPCG01N33/5082G01N2333/11C12N5/0062C12N2500/25C12N2500/84C12N2501/11C12N2513/00C12N2501/999C12N5/0688C12N2501/42C12N7/00C12N2501/30C12N2760/16011
Inventor J.周袁国勇C.李M.C.邱J.C.克莱沃斯
Owner THE UNIVERSITY OF HONG KONG