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US-20260125643-A1 - SKIN ORGANOID, METHOD FOR PRODUCING SAME, AND METHOD FOR EVALUATING DRUG BY USING SAME

US20260125643A1US 20260125643 A1US20260125643 A1US 20260125643A1US-20260125643-A1

Abstract

Provided is a method for producing a skin organoid, the method comprising: a step for performing a first culture of a primary skin cell in a first medium with at least one support among an extracellular matrix (ECM) and polyethylene glycol (PEG) to form a first skin organoid; and a step for performing a second culture of the first skin organoid in a second medium. In addition, the present disclosure provides: a skin organoid cultured by the method described above, the skin organoid having a round, spherical, three-dimensional shape and comprising a center portion, which is densely packed with cells, a stratum corneum formed on the outside of the center portion, and a hair root, wherein the cells can move from the center portion to the stratum corneum; and a method for evaluating drug toxicity by using same.

Inventors

  • Jaeyeaon CHO

Assignees

  • ORG CORP.

Dates

Publication Date
20260507
Application Date
20230927
Priority Date
20220927

Claims (20)

  1. 1 . A skin organoid comprising: a center portion (core) which is densely packed with cells; a stratum corneum formed on an outside of the center portion; and a hair root, wherein the cells are able to move from the center portion to the stratum corneum, and has a round, spherical, three-dimensional shape.
  2. 2 . The skin organoid of claim 1 , wherein the stratum corneum has a thickness of 1 to 100 μm or less.
  3. 3 . The skin organoid of claim 1 , wherein the stratum corneum includes at least one of KRT10, KRT14 and loricrin.
  4. 4 . The skin organoid of claim 1 , wherein the center portion has a diameter of 100 to 1000 μm.
  5. 5 . The skin organoid of claim 1 , wherein the center portion includes vimentin
  6. 6 . A method for producing a skin organoid including a stratum corneum formed on an outside and a hair root, the method comprising: performing a first culture of skin cells (primary skin cells) in a first medium with at least one support of an extracellular matrix (ECM) and polyethylene glycol (PEG) to form a first skin organoid; and performing a second culture of the first skin organoid in a second medium.
  7. 7 . The method of claim 6 , further comprising: before the performing a first culture, chopping a skin tissue isolated from a subject; and Obtaining heterogeneous skin cells (primary skin cells) by treating the chopped skin tissue with collagenase.
  8. 8 . The method of claim 7 , wherein the collagenase includes at least one of type I collagenase and type IV collagenase.
  9. 9 . The method of claim 7 , wherein the obtaining heterogeneous skin cells further includes culturing the primary skin cells to expand.
  10. 10 . The method of claim 6 , wherein the skin cell has a size of about 100 μm or less.
  11. 11 . The method of claim 6 , wherein the skin cell includes at least one of APOE, COL1A1, CD34, KRT7, KRT10, KRT19, HS3ST6, PECAM1 and VIM.
  12. 12 . The method of claim 6 , wherein the ECM includes at least one of a patch-shaped ECM and a fiber-shaped ECM.
  13. 13 . The method of claim 12 , wherein the fiber-shaped ECM is obtained by treating the patch-shaped ECM with a protein digestive enzyme.
  14. 14 . The method of claim 12 , wherein the fiber-shaped ECM includes at least one of collagen alpha-1(I) chain, collagen alpha-3(VI) chain, collagen alpha-2(I) chain, collagen alpha-2(VI) chain, collagen alpha-1(VI) chain, keratin type I cytoskeletal 9, keratin type II cytoskeletal 1, keratin type I cytoskeletal 10, biglycan, decorin, lumican and collagen alpha-1(III) chain.
  15. 15 . The method of claim 6 , wherein the ECM is obtained by culturing fibroblasts to form a fibroblast patch and decellularizing the fibroblast patch.
  16. 16 . The method of claim 6 , wherein the first culture is performed for at least one period of about 1 hour to 3 days.
  17. 17 . The method of claim 6 , wherein the first medium includes at least one of a B-27 supplement and a serum replacement.
  18. 18 . The method of claim 17 , wherein the B- 27 supplement is contained in an amount of 1 to 10 v/v % with respect to a total volume of the first medium.
  19. 19 . The method of claim 17 , wherein the serum replacement is contained in an amount of 5 to 15 v/v % with respect to a total volume of the first medium.
  20. 20 . The method of claim 6 , wherein the second culture is performed for at least one period of about 7 days to 40 days.

Description

TECHNICAL FIELD The present disclosure relates to a skin organoid having a structure similar to living skin, and more particularly, to a skin organoid having a round, spherical three-dimensional shape and including an outer stratum corneum, a central portion densely packed with cells, and a hair root, in which the cells move from the central portion to the stratum corneum, a method for producing the same, and a method for evaluating a drug using the same. BACKGROUND ART [Department Name] Gyeonggido Economic Science Promotion Agency [Research Project Name] Research and Development Support Project for Women Scientists and Engineers [Research Subject Name] Development of Skin Organoid Production Platform Using Human Dermal Extracellular Matrix [Research Period] Jul. 1, 2023 to Jun. 30, 2024 In the early stages of new drug development, there is a need for a model for evaluating exact toxicity and efficacy prediction. With current technology, animal models may most closely simulate the toxicity and efficacy of new drugs. However, animal experiments are burdensome in time and money, and it is difficult to completely reflect an in vivo environment of the human due to differences in genetic, biochemical and metabolic processes between species. In addition, it is difficult to technically monitor a process happening inside animals and it may also be ethically problematic. Accordingly, although primary cultured cells which are directly isolated from human tissues and cultured in vitro are used as a standard model, it is difficult to obtain tissues, and there are experimental limitations in that tissue cells do not expand in vitro. Furthermore, since a two-dimensional cell-based in vitro model is more efficient than the primary cultured cells derived from human tissues in terms of cost and labor, the model is widely used for evaluating drug toxicity and efficacy. However, the two-dimensional cell-based in vitro model is insufficient to embody the physiological functions and tissue complexity resulting from cell-cell and cell-extracellular matrix interactions. Meanwhile, an organoid is attracting attention as a new biomimetic model. The organoid is formed into a three-dimensional structure such as organs by growing stem cells into specific cells. Unlike the two-dimensional cell-based in vitro model, the organoid may be cultured in a three-dimensional environment and cultured for a longer period of time. In addition, the organoid is small in size, but constituent cells and structures thereof are similar to those of a real organ. Accordingly, the organoid has been evaluated as an optimal specimen to determine the efficacy and stability of the drug in the process of developing new drugs. Furthermore, organoid-related fields have high potential to be used not only for evaluating the drug toxicity and efficacy of new drug development, but also for disease models, cancer research, personalized medicine, regenerative therapeutics, and the like. To date, various organoids, such as stomach, intestine, early liver, thyroid, lung, brain, and the like have been successfully developed. However, a skin organoid developed to date exhibits a structurally different characteristic from a biological skin. For example, the skin organoid differs morphologically and functionally from the biological skin, as a stratum corneum and an epidermal layer are generated inside the organoid. Accordingly, a conventional skin organoid is not used as an organoid itself, but is chopped and reused as a cell-like structure, and thus has limitations in representing a biological skin in evaluation of drug and toxicity as the organoid. Therefore, there is a need to develop a skin organoid in which a stratum corneum and an epidermal layer are formed on the outside to be mimicked morphologically and functionally similar to a biological skin. The background art of the invention has been prepared to more facilitate understanding of the present disclosure. It should not be understood that the matters described in the background art of the invention exist as prior arts. DETAILED DESCRIPTION OF THE INVENTION Technical Problem More specifically, since a conventional skin organoid has a structure in which the epidermis including a stratum corneum is located inside the tissue, unlike a biological skin, the skin organoid has been used by exposing the inside of the tissue to the outside by chopping and simply layering cells. In addition, since the conventional skin organoid does not contain biological skin component cells such as vascular cells and immune cells, the skin organoid does not represent a biological skin even physiologically and functionally. Accordingly, the conventional skin organoid has been used only for a simple skin irritation test, and has not been used for evaluating material efficacy such as drug and toxicity tests and evaluating skin regeneration. Furthermore, the conventional skin organoid easily has a short culture period (expiration date) within a