Hitachi, Ltd.

Hiroshi Sonoda

Ryosuke Takahashi

Akiko Hisada

Taku Saito

Disclosed is a culture sheet which enables technology in which three-dimensional tissues with uniform diameter are formed without applying chemicals to the surface of a culture substrate. A plurality of holes are formed on the culture sheet of the substrate, and nanopillars capable of controlling the adhesiveness or migration of a cell are formed on a culture surface that serves as the bottom surface of each of the holes. The culture surface of each of the holes having a structure in which a partition wall is provided, wherein, by forming the internal nanopillars in the vicinity of the center of each of the holes, the interaction of the disseminated cells can be limited to uniform the size of the three-dimensional structures of the cells to be formed.

Culture substrate, culture sheet, and cell culture method

Provided are a technique for easily forming a spheroid by three-dimensionally culturing hepatocytes, and a technique for forming a spheroid having a higher expression level of a transporter MRP2 playing a role of biliary excretion than that of a conventional method. In order to solve the above-described problems, the present inventors have found out a condition under which hepatocytes easily form the spheroid on a nanopillar sheet. More specifically, this is related to a concentration of Type I collagen coated onto the NP sheet. Also, they have found out a condition under which an expression level of a gene related to the excretion of the formed spheroid is improved. More specifically, after the spheroid is previously formed, a biological matrix is overlayered thereon.

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Tsurugashima, Japan

Hiroshi Sonoda