Kabushiki Kaisha Toshiba

Shinshu University

Tokai National Higher Education and Research System

Daisuke Morita

Yozo Nakazawa

Eiichi Akahoshi

Mitsuko Ishihara

Yoshiyuki Takahashi

Nobuhiro Nishio

Miyuki Tanaka

According to one embodiment, there is provided a method of producing a gene modification T cell (CAR-T cells) which expresses a chimeric antigen receptor (CAR). This method includes stimulating a cell population containing a T cell with an antibody which activates the T cell, bringing the cell population into contact with a nucleic acid-introducing carrier, wherein the nucleic acid-introducing carrier containing a lipid particle, a first nucleic acid and containing a CAR gene, and a second nucleic acid containing a transposase gene encapsulated in the lipid particle, and culturing the cell population after the contacting.

Method of producing CAR-T cells, nucleic acid-introducing carrier and kit

In order to improve the efficiency of gene introduction in CAR therapy employing a transposon method, provided is a method for preparing genetically-modified T cells expressing chimeric antigen receptor, comprising: (1) a step of preparing non-proliferative cells which are obtained by stimulating a group of cells comprising T cells using an anti-CD3 antibody and an anti-CD28 antibody followed by a treatment for causing the cells to lose their proliferation capability; (2) a step of obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon method; (3) a step of mixing the non-proliferative cells prepared by step (1) with the genetically-modified T cells obtained by step (2), and co-culturing the mixed cells while stimulating the mixed cells using an anti-CD3 antibody and anti-CD28 antibody; and (4) a step of collecting the cells after culture.

Method for preparing genetically-modified T cells which express chimeric antigen receptor

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Matsumoto, Japan

Daisuke Morita