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IN VITRO MODEL FOR BLOOD-BRAIN BARRIER AND METHOD FOR PRODUCING IN VITRO MODEL FOR BLOOD-BRAIN BARRIER

Foreign code F170009280
File No. (S2016-0636-N0)
Posted date Nov 7, 2017
Country WIPO
International application number 2017JP011361
International publication number WO 2017179375
Date of international filing Mar 22, 2017
Date of international publication Oct 19, 2017
Priority data
  • P2016-081995 (Apr 15, 2016) JP
Title IN VITRO MODEL FOR BLOOD-BRAIN BARRIER AND METHOD FOR PRODUCING IN VITRO MODEL FOR BLOOD-BRAIN BARRIER
Abstract The purpose of the present invention is to provide an in vitro model in which the anatomical structure of blood-brain barrier (BBB) is reconstructed, and a method for producing an in vitro model in which the anatomical structure of BBB is reconstructed. A method for producing an in vitro model for BBB, said method comprising the following steps (a) to (e): (a) a step for culturing conditionally immortalized astrocytes on one surface of a porous membrane and culturing conditionally immortalized brain pericytes on the other surface of the porous membrane, each culture being continued until a cell sheet is formed; (b) a step for culturing conditionally immortalized brain microvascular endothelial cells in a culture container until a cell sheet is formed; (c) a step for detaching the sheet of the conditionally immortalized brain microvascular endothelial cells prepared in step (b); (d) a step for bringing the sheet of the conditionally immortalized brain microvascular endothelial cells prepared in step (c) into contact, in a layered manner, with the sheet of the conditionally immortalized brain pericytes cultured in step (a); and (e) a step for co-culturing a cell culture product prepared in step (d) that comprises three layers consisting of the sheet of the conditionally immortalized brain microvascular endothelial cells, the sheet of the conditionally immortalized brain pericytes and the sheet of the conditionally immortalized astrocytes.
Scope of claims [claim1]
1. The description below (a) the - (e) process is included, production method of blood brain channel invuitoromoderu.
(A) Conditional immortal conversion asutorosaito on the aspect of one side of the porous membrane, the process which until on the other aspect of the said porous membrane, each, it becomes seat condition, cultures conditional immortal conversion brain perisaito,
(B) In the culture container it becomes seat condition cultures the conditional the process which until, immortal conversion brain minute blood vessel endotherium,
(C) The process (b) with the process which peels the seat of the conditional immortal conversion brain minute blood vessel endotherium which is produced,
(D) The process (c) with the seat of the conditional immortal conversion brain minute blood vessel endotherium which is produced, process (a) with the process which is made to contact stratified in the seat of conditional immortal conversion brain perisaito which was cultured, and
(E) Also the cell culture which the process the process which cultures (d) with it produced, includes 3 layer of the seat of the conditional immortal conversion brain minute blood vessel endotherium, the seat of conditional immortal conversion brain perisaito and the seat of conditional immortal conversion asutorosaito
[claim2]
2. The seat and raminin layer of the conditional immortal conversion brain minute blood vessel endotherium where the aforementioned the aforementioned process (d) process (c) with produces, process (a) with in the claim 1 which features that it is the process which is made to contact stratified in the seat of conditional immortal conversion brain perisaito which was cultured production method of blood brain channel invuitoromoderu of statement. process (b) the culture surface of the culture container is done, ramininkoto with raminin layer peels the seat of the conditional immortal conversion brain minute blood vessel endotherium the aforementioned process (c) in,
[claim3]
3. The aforementioned conditional immortal conversion brain minute blood vessel endotherium, conditional immortal conversion brain perisaito and conditional immortal conversion asutorosaito, each, the first culture brain minute blood vessel endotherium, introducing the gene of the temperature sensitivity SV40 large T antigen into first culture brain perisaito and first culture asutorosaito, it features that it is something which is produced, in claim 1 or 2 production method of blood brain channel invuitoromoderu of statement.
[claim4]
4. The aforementioned process (b) in, the claim in either 1 which features that the conditional immortal conversion brain minute blood vessel endotherium is cultured in the temperature responsiveness culture container or 3 production method of blood brain channel invuitoromoderu of statement
[claim5]
5. Below from, it features that the seat of conditional immortal conversion asutorosaito, the porous membrane, in order of the seat of conditional immortal conversion brain perisaito and the seat of the conditional immortal conversion brain minute blood vessel endotherium, or, the seat of the conditional immortal conversion minute blood vessel endotherium and the seat of conditional immortal conversion brain perisaito, to the order of the seat of porous membrane and conditional immortal conversion asutorosaito it is laminated from under, blood brain channel invuitoromoderu.
[claim6]
6. Below from, it features that the seat of conditional immortal conversion asutorosaito, the porous membrane, the seat of conditional immortal conversion brain perisaito, in order of the seat of raminin layer and the conditional immortal conversion brain minute blood vessel endotherium, or, the seat of the conditional immortal conversion minute blood vessel endotherium, raminin layer, the seat of conditional immortal conversion brain perisaito, to the order of the seat of porous membrane and conditional immortal conversion asutorosaito it is laminated from under, blood brain channel invuitoromoderu.
[claim7]
7. The aforementioned conditional immortal conversion brain minute blood vessel endotherium, conditional immortal conversion brain perisaito and conditional immortal conversion asutorosaito, each, the first culture brain minute blood vessel endotherium, introducing the gene of the temperature sensitivity SV40 large T antigen into first culture brain perisaito and first culture asutorosaito, it features that it is something which is produced, in claim 5 or 6 blood brain channel invuitoromoderu of statement.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • YAMAGUCHI UNIVERSITY
  • Inventor
  • TAKESHITA YUKIO
  • KANDA TAKASHI
IPC(International Patent Classification)

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