EP-4741494-A1 - METHOD FOR CULTURING LUNG-DERIVED STEM CELLS, AND USE THEREOF

EP4741494A1EP 4741494 A1EP4741494 A1EP 4741494A1EP-4741494-A1

Abstract

Provided are a method of culturing lung-derived stem cells and use thereof, and to a method of culturing lung-derived stem cells; lung-derived stem cells prepared by the method; a pharmaceutical composition for preventing or treating hyperinflammatory diseases, the pharmaceutical composition comprising, as an active ingredient, the prepared lung-derived stem cells, a culture thereof, or cells differentiated from the lung-derived stem cells; and a quasi-drug composition. According to the culture method of the present disclosure, stem cells may be effectively isolated and obtained from lung tissues, and the obtained lung-derived stem cells have excellent pluripotency and self-renewal capacity, and excellent effects of alleviating, improving, and treating hyperinflammatory diseases, and therefore, they may be usefully applied for various purposes.

Inventors

JEON, JEONG HWA
SHIN, JI YOUNG
KIM, HEE JUNG
LEE, SEUNG JIN

Assignees

Hierabio Inc.

Dates

Publication Date: 20260513
Application Date: 20240708

Claims (15)

A method of culturing lung-derived stem cells, the method comprising the steps of: (a) embedding a lung tissue in a hydrogel and culturing the tissue to obtain a culture; and (b) decomposing the hydrogel in the obtained culture to recover stem cells that have migrated from the lung tissue into the hydrogel and proliferated therein.
The method of claim 1, wherein the step (a) comprises a pretreatment step of selecting lung tissue fragments.
The method of claim 2, wherein the pretreatment step is selecting a supernatant through centrifugation after fragmenting the lung tissue.
A lung-derived stem cell prepared by the method of claim 1.
The lung-derived stem cell of claim 4, wherein the lung-derived stem cells exhibit immunological characteristics of expressing CD90, CD29, or CD44 on the cell surface, but not expressing hematopoietic cell phenotypes CD34 and CD45.
The lung-derived stem cell of claim 4, wherein the lung-derived stem cells exhibit immunological characteristics of expressing Sonic hedgehog protein (Shh), Forkhead Box F2 (FOXF2), NK2 Homeobox 5 (Nkx-2.5), or neuroepithelial stem cell protein (Nestin) on the cell surface, but not expressing Mast/stem cell growth factor receptor kit (c-kit).
A pharmaceutical composition for preventing or treating hyperinflammatory diseases, the pharmaceutical composition comprising, as an active ingredient, the lung-derived stem cells prepared by the method of claim 1, a culture thereof, or cells differentiated from the lung-derived stem cells.
The pharmaceutical composition of claim 7, wherein the hyperinflammatory diseases are any one or more selected from respiratory inflammation, systemic inflammatory response due to respiratory inflammation, pneumonia, acute respiratory distress syndrome (ARDS), pulmonary fibrosis, cystic fibrosis, pulmonary alveolar proteinosis, lung cancer, pulmonary embolism, pulmonary edema, cytokine storm, and inflammation caused by respiratory viruses.
The pharmaceutical composition of claim 7, wherein the hyperinflammatory diseases are pneumonia or pulmonary fibrosis.
The pharmaceutical composition of claim 7, wherein the lung-derived stem cells exhibit immunological characteristics of expressing CD90, CD29, or CD44 on the cell surface, but not expressing hematopoietic cell phenotypes CD34 and CD45.
The pharmaceutical composition of claim 7, wherein the lung-derived stem cells exhibit immunological characteristics of expressing Sonic hedgehog protein (Shh), Forkhead Box F2 (FOXF2), NK2 Homeobox 5 (Nkx-2.5), or neuroepithelial stem cell protein (Nestin) on the cell surface, but not expressing Mast/stem cell growth factor receptor kit (c-kit).
A quasi-drug composition for preventing or improving hyperinflammatory diseases, the quasi-drug composition comprising, as an active ingredient, the lung-derived stem cells prepared by the method of claim 1, a culture thereof, or cells differentiated from the lung-derived stem cells.
A composition for culturing the lung-derived stem cells of claim 1, the composition comprising a lung tissue and a hydrogel.
A kit for culturing lung-derived stem cells, the kit comprising the composition for culturing stem cells of claim 13.
Use of a composition comprising lung-derived stem cells prepared by the method of claim 1, a culture thereof, or cells differentiated from the lung-derived stem cells as an active ingredient in preventing, treating, or improving hyperinflammatory diseases.

Description

[Technical Field] The present disclosure relates to a method of culturing lung-derived stem cells and use thereof, and to a method of culturing lung-derived stem cells; lung-derived stem cells prepared by the method; a pharmaceutical composition for preventing or treating hyperinflammatory diseases, the pharmaceutical composition comprising, as an active ingredient, the prepared lung-derived stem cells, a culture thereof, or cells differentiated from the lung-derived stem cells; and a quasi-drug composition. [Background Art] Stem cells refer to cells in a pre-differentiation stage before they differentiate into individual cells that make up a tissue, and to cells that can proliferate infinitely in an undifferentiated state and have the potential to differentiate into various types of tissue cells through specific differentiation stimulation. According to their differentiation potency, stem cells are largely divided into embryonic stem cells (ES cells) and adult stem cells (tissue-specific stem cells). Embryonic stem cells are stem cells isolated from the inner cell mass (ICM), which will develop into a fetus, of a blastocyst embryo, the initial stage in which the fertilized egg is formed but is not yet implanted in the endometrium, and are cells that have the potential to differentiate into cells of all tissues. In contrast, tissue-specific stem cells are stem cells specific to each organ that appear in the stage in which the embryonic development process progresses and each organ of the embryo is formed, and their differentiation potency is generally limited (multipotent) only to the cells that make up the tissue. Representative tissue-specific stem cells comprise hematopoietic stem cells, which exist in the bone marrow, and mesenchymal stem cells, which differentiate into connective tissue cells other than blood cells. Hematopoietic stem cells differentiate into various blood cells such as erythrocytes, leukocytes, etc., and mesenchymal stem cells differentiate into osteoblasts, chondroblasts, adipocytes, and myoblasts, etc. Recently, since the success of the isolation of embryonic stem cells from humans, interest in their clinical application has grown. The application of stem cells that is receiving the most attention is the use of stem cells as a cell source for cell replacement therapy. The technology of isolating/culturing lung-derived stem cells of the present disclosure is a technology utilizing a hydrogel-based niche-preserving, self-renewal-induced isolation technology for stem cell isolation, and is a novel technology that enables the ex vivo survival/culture of lung tissue and the direct isolation of highly pure cells from the tissue without enzymatic treatment. Existing methods of isolating lung-derived stem cells rely on enzymatic isolation methods using collagenase, etc. (Korean Patent Publication No. 10-2014-0075469 A), and these methods inevitably damage cell surface proteins, affecting cell performance. However, the present isolation method is distinct from the existing methods, and has an advantage of being superior in terms of ex vivo expansion and cell performance. [Disclosure] [Technical Problem] Cells isolated using existing methods are able to be expanded ex vivo only for 2 passages to 3 passages (up to 5 passages). However, cells isolated using a method of the present disclosure exhibit superior performance, such as no cell senescence even after passages of 8 or more. Therefore, long-term passage culture is possible for mass production. Under this background, the present inventors have conducted extensive research to develop novel stem cells. As a result, they found that lung-derived stem cells prepared by the culture method of the present disclosure exhibit superior ex vivo expansion, cell performance, and differentiation potency, and accordingly, they may be used for the treatment of hyperinflammatory diseases, thereby completing the present disclosure. [Technical Solution] An object of the present disclosure is to provide a method of culturing lung-derived stem cells using a lung tissue and a hydrogel. Another object of the present disclosure is to provide lung-derived stem cells prepared by the above method. Still another object of the present disclosure is to provide a pharmaceutical composition for preventing or treating hyperinflammatory diseases, the pharmaceutical composition comprising, as an active ingredient, the lung-derived stem cells, a culture thereof, or cells differentiated from the lung-derived stem cells. Still another object of the present disclosure is to provide a quasi-drug composition for preventing or improving hyperinflammatory diseases, the quasi-drug composition comprising, as an active ingredient, the lung-derived stem cells, a culture thereof, or cells differentiated from the lung-derived stem cells. Still another object of the present disclosure is to provide a composition for culturing lung-derived stem cells, the composition comprising a lung tissu