Cell culture substrate, cancer cell aggregate and method for manufacturing same using said substrate, and drug screening method using said cancer cell aggregate
[Problem] An object of the present invention is to produce cancer cell clusters with intrinsic biological properties as cancer tissues, such as morphological polarity and tissue motion polarity, in vitro. [Solution] The present invention relates to a cell culture substrate including a base material and a biocompatible polymer layer, the substrate including a plurality of rough sections on the surface of the substrate, wherein the rough sections are not covered with the biocompatible polymer layer, have a predetermined surface structure with a predetermined shape, and are disposed at predetermined intervals. With the present invention, it is possible to obtain a live cancer cell aggregate having morphological polarity and tissue motion polarity similar to that observed in vivo, by a very easy operation of culturing cancer cells on a cell culture substrate having a predetermined structure, thereby performing live imaging of microtumors in vitro is enabled, which has been conventionally impossible. Moreover, since such a cancer cell aggregate is considered to reproduce a series of flow of development, proliferation, infiltration, metastasis, and recurrence of cancer in vivo, the cancer cell aggregate can be utilized as a research tool in cancer research, or for screening for an anticancer drug.
1. A cell culture substrate, comprising:
a base material having a plurality of rough sections; and
a biocompatible polymer layer covering the base material such that the plurality of rough sections is exposed on a surface of the cell culture substrate,
wherein
each of the rough sections has a shape of a spot with a diameter ranging from 20 μm to 100 μm, or a groove with a width ranging from 3 μm to 30 μm, when the shape of the rough section is a groove, an end part of the rough section is optionally connected to another rough section,
a distance between two adjacent rough sections is at least 10 μm or more, and the rough section has at least one uneven structure with a height ranging from 20 nm to 200 nm on the surface.
2. The cell culture substrate according to claim 1 , wherein the rough section has a developed interfacial area ratio (Sdr) of 0.002 or more.
3. The cell culture substrate according to claim 1 , wherein the distance between the two adjacent rough sections ranges from 10 to 1,200 μm.
4. The cell culture substrate according to claim 1 , wherein the rough section has an arithmetic mean roughness (Ra) of 4 nm or more, a maximum height roughness (Rz) of 30 nm or more, and/or an arithmetic mean peak curvature (Spc) of 300 or more.
5. The cell culture substrate according to claim 1 , wherein the biocompatible polymer is an amphiphilic polymer that inhibits non-specific adsorption to a biological material.
6. The cell culture substrate according to claim 5 , wherein the amphiphilic polymer is 2-methacryloyloxyethyl phosphorylcholine.
7. The cell culture substrate according to claim 5 , wherein the amphiphilic polymer is selected from the group consisting of dimethyl polysiloxane, polyethylene glycol, oligoethylene glycol and 2-methacryloyloxyethyl phosphorylcholine.
8. The cell culture substrate according to claim 1 , wherein the rough section has an arithmetic mean roughness (Ra) of 4 nm or more.
9. The cell culture substrate according to claim 1 , wherein the rough section has a maximum height roughness (Rz) of 30 nm or more.
10. The cell culture substrate according to claim 1 , wherein the rough section has an arithmetic mean peak curvature (Spc) of 300 or more.
11. The cell culture substrate according to claim 1 , wherein the height of the uneven structure ranges from 30 nm to 100 nm.
12. The cell culture substrate according to claim 1 , wherein the height of the uneven structure ranges from 40 nm to 60 nm.