JP-2026076380-A - Gel composition, method for manufacturing the same, and three-dimensional structure, method for manufacturing the same

JP2026076380AJP 2026076380 AJP2026076380 AJP 2026076380AJP-2026076380-A

Abstract

[Problem] To provide a gel composition containing extracellular matrix components and/or fragmented extracellular matrix components that is transparent and has a high elastic modulus. [Solution] A gel composition containing at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components, and ions of a metal element. [Selection Diagram] None

Inventors

北野史朗
松▲崎▼ 典弥
櫻井英博
植竹裕太

Assignees

ＴＯＰＰＡＮホールディングス株式会社
国立大学法人大阪大学

Dates

Publication Date: 20260511
Application Date: 20260224
Priority Date: 20200622

Claims (12)

It contains at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components, and an ion of a metal element. The extracellular matrix component includes collagen, The aforementioned fragmented extracellular matrix component includes defibrated collagen, The aforementioned metallic element is at least one metallic element selected from the group consisting of transition metals of Group 10, transition metals of Group 11, and metals of Group 12 of the periodic table. A gel composition having an elastic modulus of 1.5 kPa or higher.
The gel composition according to claim 1, wherein the elastic modulus is 1686 kPa or less.
The gel composition according to claim 1 or 2, wherein the aforementioned metal element is at least one selected from the group consisting of copper, zinc, palladium, platinum, and gold.
The gel composition according to any one of claims 1 to 3, wherein at least one selected from the group consisting of the extracellular matrix component and the fragmented extracellular matrix component is crosslinked.
A method for producing a gel composition, The method comprises a step of contacting a solution containing at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components with a solution containing ions of a metal element. The extracellular matrix component includes collagen, The aforementioned fragmented extracellular matrix component includes defibrated collagen, The aforementioned metallic element is at least one metallic element selected from the group consisting of transition metals of Group 10, transition metals of Group 11, and metals of Group 12 of the periodic table. A method for producing the gel composition, wherein the elastic modulus of the gel composition is 1.5 kPa or more.
The manufacturing method according to claim 5, wherein the content of at least one selected from the group consisting of the extracellular matrix component and the fragmented extracellular matrix component in the gel composition is 0.01% by weight or more and 1% by weight or less, based on the total amount of the gel composition.
The manufacturing method according to claim 5 or 6, wherein the content of the metal element ions in the gel composition is 5 mg to 60 mg per 1 g of the total content of at least one selected from the group consisting of the extracellular matrix component and the fragmented extracellular matrix component.
A step of obtaining a cell-containing gel composition by contacting at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components, cells, and ions of a metal element in an aqueous medium. The process comprises a step of culturing the cell-containing gel composition, The extracellular matrix component includes collagen, The aforementioned fragmented extracellular matrix component includes defibrated collagen, The aforementioned metallic element is at least one metallic element selected from the group consisting of transition metals of Group 10, transition metals of Group 11, and metals of Group 12 of the periodic table. A method for producing a three-dimensional structure, wherein the elastic modulus of the gel composition is 1.5 kPa or more.
The method for producing a three-dimensional tissue according to claim 8, comprising the step of obtaining the cell-containing gel composition, wherein the step of contacting a suspension containing at least one selected from the group consisting of the extracellular matrix component and fragmented extracellular matrix component, the cells, and a first aqueous medium with a solution containing the ions of the metal element and a second aqueous medium.
The method for producing a three-dimensional tissue according to claim 8 or 9, wherein the culture step is performed under conditions that allow at least a portion of the cells to maintain a viable state.
A method for producing a three-dimensional tissue according to any one of claims 8 to 10, further comprising the step of incubating the cell-containing gel composition at 20°C to 30°C after the step of obtaining the cell-containing gel composition and before the step of culturing.
It contains at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components, cells, and ions of a metal element. The extracellular matrix component includes collagen, The aforementioned fragmented extracellular matrix component includes defibrated collagen, The aforementioned metal element is at least one metal element selected from the group consisting of transition metals of Group 10, transition metals of Group 11, and metals of Group 12 of the periodic table, and has a total light transmittance of 80% or more at 37°C. A three-dimensional tissue with a thickness of 10 μm or more.

Description

This invention relates to a gel composition and a method for producing the same. The invention also relates to a three-dimensional structure using the gel composition according to the present invention, and a method for producing the same. Gels of extracellular matrix components such as collagen are commonly used as scaffold materials in cell culture. Collagen gels are typically prepared by dissolving collagen under acidic conditions, followed by neutralization and heating to approximately 37°C to gel the collagen. Another known method involves cross-linking collagen using cross-linking agents such as formaldehyde and glutaraldehyde. On the other hand, for example, Patent Document 1 discloses an extracellular matrix-containing composition comprising fragmented extracellular matrix components and an aqueous medium. Since the extracellular matrix-containing composition disclosed in Patent Document 1 exhibits a thermally reversible sol-gel transition, gelation and solification can be controlled by temperature. International Publication No. 2019/208831 This is a photograph showing the results of Test Example 1.This is a photograph showing the results of Test Example 2.This is a photograph showing the results of Test Example 3.This is a photograph showing the results of Test Example 4.These are photographs showing the external appearance (Figure 5(A)) and internal structure (Figure 5(B)) of the three-dimensional tissue produced in Test Example 6.This graph shows the elastic modulus of gel compositions prepared using solutions of various metal ions.(A) A graph showing the transmittance at a wavelength of 500 nm for gel compositions prepared with fragmented collagen solution and Pt2 + solution at various concentrations. (B) Images of the gels prepared with fragmented collagen solution and Pt2 + solution at various concentrations, observed from above.This is a photograph showing the results of Test Example 9. The following describes in detail embodiments for carrying out the present invention. However, the present invention is not limited to the following embodiments. [Gel composition] The gel composition according to this embodiment contains at least one selected from the group consisting of extracellular matrix components and fragmented extracellular matrix components, and ions of a metal element. Extracellular matrix components are aggregates of extracellular matrix molecules, formed by multiple extracellular matrix molecules. Extracellular matrix molecules may be any substances present outside the cell in a multicellular organism. Any substance can be used as an extracellular matrix molecule, as long as it does not adversely affect cell growth or the formation of cell aggregates. Examples of extracellular matrix molecules include, but are not limited to, collagen, laminin, fibronectin, vitronectin, elastin, tenascin, entericin, fibrillin, and proteoglycans. These extracellular matrix molecules may be used individually or in combination of two or more. The extracellular matrix component may, for example, contain collagen, or may consist solely of collagen. When the extracellular matrix component contains collagen, the effects of the present invention—that the gel composition is transparent and has a high elastic modulus (rigidity)—can be more pronounced. Furthermore, when the extracellular matrix component contains collagen, and the gel composition according to this embodiment is used as a scaffold material during cell culture, the collagen functions as a scaffold for cell adhesion, further promoting the formation of three-dimensional cell structures. The extracellular matrix molecule may be a modified or variant of the extracellular matrix molecule described above, or it may be a polypeptide such as a chemically synthesized peptide. The extracellular matrix molecule may have a repeating sequence represented by Gly-X-Y, which is characteristic of collagen. Here, Gly represents a glycine residue, and X and Y each independently represent any amino acid residue. Multiple Gly-X-Y sequences may be identical or different. Having a repeating sequence represented by Gly-X-Y reduces constraints on the arrangement of the molecular chain, resulting in, for example, even better functionality as a scaffold material during cell culture. In an extracellular matrix molecule having a repeating sequence represented by Gly-X-Y, the proportion of the sequence represented by Gly-X-Y may be 80% or more of the total amino acid sequence, preferably 95% or more. The extracellular matrix molecule may also be a polypeptide having an RGD sequence. An RGD sequence refers to a sequence represented as Arg-Gly-Asp (arginine residue-glycine residue-aspartic acid residue). The presence of an RGD sequence further promotes cell adhesion, making it more suitable, for example, as a scaffold material in cell culture. Examples of extracellular matrix molecules containing both a Gly-X-Y sequence and an RGD sequence include collagen, fibronectin, vitron