JPWO2019152409A5 - - Google Patents

Claims (21)

A method of increasing the therapeutic efficacy of mammalian cells suitable for use in cell therapy, wherein the less potent or slightly more potent mammalian cells are brought into contact with an extrinsic agent to sufficiently bring the mammalian cells into contact. The mammalian cell comprises the step of increasing the expression level of the target transcription factor in the cell to the extent that it transforms into a potent tissue effector cell , thereby producing an activation-inducing tissue effector cell (AITEC). Is a method comprising Cardiosphere-derived cells (CDCs) or fibroblasts . A method of inducing activation of weak or not strong enough mammalian cells suitable for use in cell therapy, wherein the method is a sufficiently strong tissue effector for the weak or not strong enough cells. Including the step of introducing an exogenous agent that increases the level of the target transcription factor in the cell to the extent of conversion into the cell into the weak or not sufficiently strong cell , thereby producing AITEC. The method, wherein the mammalian cell comprises a cardiosphere-derived cell (CDC) or a fibroblast . The method of claim 1 or 2 , further comprising the step of selecting the AITEC for use in cell therapy. The steps in selecting AITEC for use in cell therapy include inclusion of the selected gene within the gene delivery system, treatment of the cells with antibiotics, and cells that do not successfully deliver the gene delivery system. The method according to claim 3 , which is carried out by thinning out. The method according to any one of claims 1 to 4 , wherein the target transcription factor is β-catenin. The method according to any one of claims 1 to 5 , wherein the mammalian cell is a human cell. The method of claim 6 , wherein the human cell is a weak cardiosphere-derived cell (CDC). The method of claim 7 , wherein the CDC is an immortalized CDC. The immortalized CDC
(A) A step of overexpressing the small T antigen and the large T antigen of SV40 in the culture medium of CDC, and
(B) The method of claim 8 , wherein the method is produced by a method comprising selecting a CDC culture medium capable of continuing the doubling at least 15 times. The immortalized CDC
(A) A step of overexpressing c-Myc in the culture medium of CDC and
(B) The method of claim 8 , wherein the method is produced by a method comprising selecting a CDC culture medium capable of continuing the doubling at least 15 times. The method according to any one of claims 1 to 10 , wherein the mammalian cells are plate-cultured in a fibronectin-coated culture vessel. The method according to any one of claims 1 to 11 , further comprising increasing the level of another target transcription factor in the mammalian cell. 12. The method of claim 12 , wherein the other target transcription factor is GATA4. Any one of claims 1-13 , wherein the levels of β-catenin in non-strong cells are increased by administering 6-bromoinsylbin-3'-oxime (BIO), Wnt3a, or CHIR to the mammalian cells. The method described in the section . AITEC produced by the method according to any one of claims 1 to 14 . A pharmaceutical composition comprising a therapeutically effective amount of AITEC-derived extracellular vesicles produced by the method according to any one of claims 1 to 14 . The pharmaceutical composition according to claim 16 , wherein the extracellular vesicle is an exosome. A pharmaceutical composition comprising a therapeutically effective amount of AITEC according to claim 15 for treating a disease or condition in a subject in need thereof. A pharmaceutical composition comprising a therapeutically effective amount of AITEC according to claim 15 , for treating a subject in need of cell therapy for treating a disease or condition by administering the AITEC to a target cell. .. 19. The pharmaceutical composition of claim 19 , wherein administration of AITEC induces activation of the β-catenin / Wnt pathway in the target cells. 20. The pharmaceutical composition of claim 20 , wherein administration of AITEC induces activation of at least one of anti-inflammatory, regeneration, attenuation of fibrosis, and angiogenesis.