JP-2022549245-A5 -

Dates

Publication Date

20230511

Application Date

20200710

Description

Endothelial cell production using various other protocols, such as PSC differentiation and direct reprogramming of somatic cells, has not been reported to achieve a sufficient number of functional endothelial cells within the timeframe desired for clinical application. A major difficulty in the clinical use of reprogrammed endothelial cells (ECs) is the low maintenance efficiency of ECs injected into the patient's ischemic target area. The method disclosed herein has several advantages compared to other EC production methods. Source cells can be supplied without limitation from urine using non-invasive methods. EC production can be performed more timely compared to other methods. Reprogrammed endothelial and smooth muscle cells can be obtained together in cell sheet structures that maintain well in ischemic areas without the use of heterologous biomaterials. The reprogrammed vascular-like tissue is applicable to autologous cell therapy for various ischemic diseases such as PAD and MI. Embodiments of the present invention Aspects of the present invention are further described in the following sections: [Section 1] i) Concentrating urinary cells from the subject; ii) The cells of the concentrated urine a) EGF, b) Hydrocortisone, c) Epinephrine, and d) Human serum or animal serum To provide purified concentrated urine-derived cells by replicating in a first growth medium containing [a specific substance]; iii) Exposing the purified concentrated urine-derived cells to ETV2; iv) To provide endothelial-like urine-derived cells by culturing the purified concentrated urine-derived cells in a first growth medium; v) The endothelial urine-derived cells a) EGF, b) VEGFA, c) bFGF, d) Heparin, e) L-ascorbic acid, and d) Human serum or animal serum To provide endothelial and smooth muscle-like vascular tissue by culturing in a second growth medium containing [the specified substance]. A method for producing endothelial and smooth muscle-like vascular tissue, including [the specified element]. [Section 2] The method described in item 1 above, wherein human serum is derived from the subject. [Section 3] The method according to item 1, wherein the purified concentrated urine-derived cells are exposed to ETV2, and the purified concentrated urine-derived cells are mixed with a recombinant virus that infects the purified concentrated urine-derived cells, contains a gene encoding ETV2, and expresses ETV2 after infection. [Section 4] The method according to item 3 above, wherein the recombinant virus is an adenovirus or a lentivirus. [Section 5] The method according to item 1, wherein the growth medium comprises glucose, amino acids, and vitamins, glutamine, and sodium pyruvate. [Section 6] The method according to item 1, further comprising the step of folding the endothelial and smooth muscle-like vascular tissue into a three-dimensional structure. [Section 7] The method according to item 1, further comprising transplanting endothelial and smooth muscle-like vascular tissue. [Section 8] The method according to item 7, wherein transplantation of endothelial and smooth muscle-like vascular tissue is performed by bringing the endothelial and smooth muscle-like vascular tissue into contact with veins, arteries, capillaries, myocardium, skin, lungs, kidneys, or intestines.