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TECHNIQUE FOR AGGREGATING MACROMOLECULES TOGETHER WITH CELLS

Foreign code F170008919
File No. (S2015-1519-N0)
Posted date Jan 19, 2017
Country WIPO
International application number 2016JP064167
International publication number WO 2016182022
Date of international filing May 12, 2016
Date of international publication Nov 17, 2016
Priority data
  • P2015-099064 (May 14, 2015) JP
Title TECHNIQUE FOR AGGREGATING MACROMOLECULES TOGETHER WITH CELLS
Abstract Provided is a technique for efficiently aggregating macromolecules such as ECM together with cells. A method for creating a cell or a cell aggregate loaded with macromolecules, the method comprising adding, to a medium containing a swellable material, a solution containing macromolecules and at least one cell to aggregate the macromolecules together with the cell. A method for controlling the function and/or the property of a cell or a cell aggregate, the method comprising culturing the cell or the cell aggregate loaded with the macromolecules created in the aforementioned method. A method for culturing a cell or a cell aggregate, the method comprising: adding, to a medium containing a swellable material, a solution containing macromolecules and at least one cell to aggregate the macromolecules together with the cell and to produce a macromolecule capsule encapsulating the cell or the cell aggregate; and culturing the cell or the cell aggregate in the capsule. For preparing a three-dimensional cell tissue, methods such as 96-well U-bottom plating and hanging drop technique are conventionally used. In the methods of the present invention, 2000 cells are mixed with 1 μl of the medium containing ECM, and the ECM and cells can be simultaneously aggregated. If the efficiency here is defined as 1, a calculation can be made that, when 96-well U-bottom plating or hanging drop technique is used, ECM can only be mixed in the three-dimensional cell tissue at an efficiency of about 1/12000 or about 1/2400, respectively. Thus, with the present invention, the cost for ECM can be reduced to about one several-thousandth to one hundred-thousandth, and it becomes possible to effectively use artificial ECM that cannot be synthesized in large quantity or rare ECM that can be extracted only at a very small amount because of reasons such as the subject being a small living organism.
Outline of related art and contending technology BACKGROUND ART
Cells were used for cell assays or in regenerative medicine, cells were three-dimensional organization of cells in order to enhance the ability to differentiate is considered to be an effective means. Is a general three-dimensional organization, cell clumps in order to make the non-adhesive plate or the hanging drop method using (Lin et al., Biotechnology Journal, 2008,3,1172-1184: non-patent document 1). However, as is required for the regenerative medicine and in drug screening, with high added value than for production of three-dimensional tissue, which further devised method for constructing a three-dimensional tissue is demanded.
We present in the body of the organs, cells and extracellular matrix and laminated Mn. Previously and in the extracellular matrix (ECM) on the cell surface which is a kind of gelatin and fibronectin coating, laminating technique (Nishiguchi et al., Advanced Materials, 2011, 23, 3506-3510: non-patent document 2, Matsusaki et al., Angewandte Chemie 2007, 46, 4689-4692: non-patent document 3) is reported.
Scope of claims (In Japanese)[請求項1]
膨潤性材料を含有する培地へ高分子と少なくとも1個の細胞とを含む溶液を添加して、高分子と細胞を共に凝集させることを含む、高分子が充填された細胞又は細胞凝集体を作製する方法。
[請求項2]
細胞凝集体が、3次元組織を形成する請求項1記載の方法。
[請求項3]
3次元組織の性状及び/又は機能が、3次元組織を形成していない細胞の性状及び/又は機能又は細胞間に高分子を充填せずに形成された3次元組織の性状及び/又は機能より向上している請求項2記載の方法。
[請求項4]
高分子が充填された細胞又は細胞凝集体が、細胞又は細胞凝集体を封入した高分子のカプセルである請求項1~3のいずれかに記載の方法。
[請求項5]
膨潤性材料を含有する培地へ添加する溶液に含まれる少なくとも1個の細胞が、請求項1~4のいずれかに記載の方法で作製した高分子が充填された細胞又は細胞凝集体である請求項1~4のいずれかに記載の方法。
[請求項6]
請求項1~5のいずれかに記載の方法で作製した、高分子が充填された細胞又は細胞凝集体を培養することを含む、細胞又は細胞凝集体の性状及び又は機能を制御する方法。
[請求項7]
膨潤性材料を含有する培地へ高分子と少なくとも1個の細胞とを含む溶液を添加して、高分子と細胞を共に凝集させることで、細胞又は細胞凝集体を封入した高分子のカプセルを作製し、このカプセル内で細胞又は細胞凝集体を培養することを含む、細胞又は細胞凝集体の培養方法。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • PUBLIC UNIVERSITY CORPORATION YOKOHAMA CITY UNIVERSITY
  • Inventor
  • KOJIMA, Nobuhiko
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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