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Method for providing vascular system in biological tissue

A vascular system and biological technology, applied in the field of making three-dimensional tissue with vascular network, can solve the problems of independent stent material, unestablished, foreign body reaction of stent material, etc., and achieve the effect of high technical value

Active Publication Date: 2016-03-02
PUBLIC UNIV CORP YOKOHAMA CITY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, when used in transplantation, etc., the foreign body reaction of the stent material, causing inflammation, etc. is also considered to be a big problem
[0006] As mentioned above, although the reconstruction of a three-dimensional tissue with a vascular network is desired for presumed industrial applications or regenerative medicine applications, the current situation is that a method for constructing an organoid with blood vessels using tissue in vitro has not yet been established.

Method used

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  • Method for providing vascular system in biological tissue
  • Method for providing vascular system in biological tissue
  • Method for providing vascular system in biological tissue

Examples

Experimental program
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Effect test

Embodiment approach

[0084] Hereinafter, the present invention will be described in detail.

[0085] The present invention provides a method for imparting a vascular system to a biological tissue in vitro, which comprises co-culturing the biological tissue with vascular cells and mesenchymal cells.

[0086] In this specification, "biological tissue" refers to a structure composed of a plurality of cells, and examples include normal-abnormal tissue or cancer tissue isolated from an individual, self-pluripotent stem cells (artificial pluripotent stem cells (iPS cells) , embryonic stem cells (ES cells), etc.), tissue stem-precursor cells, differentiated cells, etc. induced tissues), etc. Biological tissues can be mainly of human origin, but animals other than humans (e.g., animals used for laboratory animals, companion animals, working animals, racehorses, fighting dogs, etc., specifically, mice, rats, Biological tissues derived from rabbit, pig, dog, monkey, cow, horse, sheep, chicken, shark, ray, ...

Embodiment 1

[0130] [Example 1: Implantation of vascular network in islet tissue]

[0131] 〔method〕

[0132] 【1. Isolation method of mouse islets】

[0133] Isolation of mouse islets was mainly carried out according to the method of Dong et al. (document name: Aprotocolforisletisolationfrommousepancreas). The abdomen of an anesthetized C57BL / 6J mouse (Japan SLC) was sterilized with 70% ethanol using diethyl ether (Wako), and the abdomen was opened, and the ampulla of Farter, which is the junction of the common bile duct and the duodenum, was ligated. Thereafter, a 27G injection needle was inserted into the confluence of the cystic duct and the hepatic duct, and 3 ml of collagenase XI solution (1,000 U / ml) (Sigma, cat. No. C7657) prepared with Hanks buffer (HBSS, GIBCO) was injected, Fill the whole pancreas with collagenase XI solution. Cut out the pancreas, put it into a 50ml tube containing collagenase XI solution, and shake it at 37.5°C for 15 minutes. After digesting the pancreas, ad...

Embodiment 2

[0215] [Example 2: Delivery of vascular network to renal glomeruli]

[0216] 〔method〕

[0217] 【1. Isolation method of mouse glomeruli】

[0218] After the abdomen of a C57BL / 6-Tg mouse (Japan SLC) anesthetized with diethyl ether (Wako) was disinfected with 70% ethanol, the abdomen was opened, and the kidney was removed. The renal epithelium was removed from the excised kidney, and after washing with physiological saline, the kidney was cut with a scalpel wheel. The renal pelvis and medulla were removed with clips, and the cortex was recovered. The recovered cortex was minced on ice, and filtered by adding Hanks buffer (HBSS, GIBCO) containing 0.1% bovine serum albumin (BSA, SIGMA) little by little using a 100 μm mesh cell strainer. The flow-through was filtered through a 70 μm mesh cell filter, and finally, the flow-through was filtered through a 40 μm mesh cell filter. The cell mass remaining on the 40 μm purpose cell filter was collected using Hanks buffer containing 0.1...

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Abstract

The invention provides a method for providing a vascular system in biological tissue in vitro. The method comprises cocultivation of the biological tissue with vascular cells and mesenchymal cells. Biological tissue in which a vascular system has been provided by said method. A method for producing a tissue or organ, the method comprising transplantation of the biological tissue in a non-human animal and inducing differentiation to a tissue or organ in which a vascular network has been constructed. A tissue or organ regeneration or function-restoring method comprising transplantation of the biological tissue in a human or non-human animal and inducing differentiation to a tissue or organ in which a vascular network has been constructed. A method for producing a non-human chimera animal, the method comprising transplantation of the biological tissue to a non-human animal and inducing differentiation to a tissue or organ in which a vascular network has been constructed. A method for evaluating drugs using at least one selected from a group consisting of the biological tissue, tissues and organs produced by said method, and non-human chimera animals produced by said method. A regenerative therapy composition comprising biological tissue in which a vascular system has been provided by the method.

Description

【Technical field】 [0001] The present invention relates to a method for imparting a vascular system to a biological tissue, and more specifically relates to a method for producing a three-dimensional tissue with a vascular network from a tissue induced from pluripotent stem cells, etc., or from a tissue such as normal tissue or cancer tissue isolated from an individual method. 【Background technique】 [0002] In recent years, attention has been paid to regenerative medicine that utilizes normal / cancer tissues isolated from individuals or tissues induced from pluripotent stem cells to realize drug development screening for the development of new drugs or to compensate for functions of missing organs. [0003] As an attempt to induce three-dimensional tissues from pluripotent stem cells, etc., it has been reported that small spherical tissues are formed in areas such as the liver, pancreas, and nerves, and cell differentiation is induced (Non-Patent Document 1: Takayama K, et al...

Claims

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Application Information

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IPC IPC(8): C12N5/071A01K67/027A61L27/00C07D499/21G01N33/50
CPCC07D499/21G01N33/5082A61L27/3808A61L27/3834A61L27/3886A01K67/0271A01K2207/12A01K2267/0325A01K2267/0331A01K2267/035A01K2267/0362C12N5/0691A01K2227/105C12N5/0062C12N5/0677C12N2506/1346C12N2506/28C12N2513/00A61K35/28A61K35/44A61K49/0008C12N2502/13
Inventor 武部贵则谷口英树高桥祯畅
Owner PUBLIC UNIV CORP YOKOHAMA CITY UNIV
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