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CELL SHEET FOR VITAL TRANSPLANTATION AND METHOD FOR PRODUCING SAME

Foreign code F200010008
File No. (17004-JP01)
Posted date Jan 29, 2020
Country WIPO
International application number 2019JP017594
International publication number WO 2019208688
Date of international filing Apr 25, 2019
Date of international publication Oct 31, 2019
Priority data
  • P2018-084565 (Apr 25, 2018) JP
Title CELL SHEET FOR VITAL TRANSPLANTATION AND METHOD FOR PRODUCING SAME
Abstract The present invention pertains to a cell sheet for vital transplantation, said cell sheet being provided with mesenchymal stem cells on a surface at an average cell density of 3.0×104 cells/cm2 or less. The present invention also pertains to a method for producing a cell sheet for vital transplantation, said method comprising a step for inoculating mesenchymal stem cells in a cell culture support having a three-dimensional structure formed of a fiber at a ratio of 3.0×105 cells/cm2 or less and a step for culturing the mesenchymal stem cells to thereby prepare a cell sheet having an average cell density of 3.0×104 cells/cm2 or less.
Outline of related art and contending technology BACKGROUND ART
Mesenchymal stem cells (Mesenchymal stem cell, hereinafter also referred to as MSC) is, perhaps the ability to self-replicating and have the ability to a stem cell, osteoblast, chondrocyte cells, fat cells, muscle cells to differentiate into cells such as mesenchymal belonging to the well, such as hepatocytes or cells also have the ability to differentiate than definitive. In the MSC, itself produced by the liquid agent and cell adhesion interactions paracrine effect have been known. MSC is, based on these actions, the repair of the target tissue or cell, a regenerative capacity, and exhibits an anti-inflammatory such as ability to control, as a result, various diseases are considered to represent the therapeutic effect.
The MSC, can be easily isolated and cultured up the force in the growth, the number of cells in a short time can be secured to the implantable, immune rejection can be autografts can not, be less ethical issues, it is not necessary to pre-treatment by low immunogenicity allograft practical reasons, as the material of the ideal cell transplantation therapy, a treatment for various diseases has been applied to study.
MSC of the cell transplantation therapy of disease expected to be applied to one of the renal disease, particularly chronic kidney disease. Chronic kidney disease are, renal disorders represented by proteinuria, and glomerular filtration rate as an indicator of renal function either or both of the state has persisted for more than six months refers to 3, 8 adult person in Japan and affecting 1. Chronic kidney disease symptoms and progress to end stage renal disease, drug treatment effect cannot be expected, an increase in the artificial dialysis in many patients. Artificial dialysis be symptomatic, chronic renal failure patients due to a need to continue to receive an artificial dialysis over the lifetime, the physical and economic burden to a patient is large. Further increase in dialysis medical costs, medical economy, a concern has been a problem.
The inventors of the present invention, the MSC is used to establish cell transplantation therapy in the course of, for example a diabetic patient in the patient's abnormal MSC in the MSC, specifically as described above or a variety of capability of the capability of the MSC and compared with that of a normal as a result, the effect of disease or therapeutic effect is lost as compared with the normal MSC MSC is reduced, and mammals extract from the associated MSC and activating the abnormal treatment efficacy can be restored, and such activation treatment using autografts the MSC can be found and the like, attached to mammals as an active ingredient extract from abnormal activation agent for the MSC to the invention, and filed a patent application (Patent Document 1). The activation agent is, in particular, the treatment step becomes necessary for the patient at a point of the MSC can be autografts, has important significance.
On the other hand, in the cell transplantation therapy, used to reduce the amount of the MSC, centralized MSC sites required to be capable of formulation of the topical application is being examined. Of the cell transplantation therapy to renal disease as the MSC, the MSC of the sheet composition is applied to the kidney cell can approach is to treat kidney disease have been reported (Patent Document 2). The cells of the sheet composition is a stimuli-sensitive media on the substrate and seeded culture MSC, to separate the cells stimulated by peeling are prepared.
Scope of claims (In Japanese)[請求項1]
 平均細胞密度が3.0×10 4細胞/cm 2以下の間葉系幹細胞をその表面に有する、生体移植用細胞シート。

[請求項2]
 さらに生体適合性の支持体を含む、請求項1に記載の細胞シート。

[請求項3]
 支持体がファイバーからなる3次元構造を有する細胞培養担体である、請求項2に記載の細胞シート。

[請求項4]
 細胞培養担体が、ナノメートル~マイクロメートル単位の平均繊維径を有するファイバーによって細胞との接触面上に形成された開口部を有する、請求項3に記載の細胞シート。

[請求項5]
 開口部の平均径が500nm~1000μmである、請求項4に記載の細胞シート。

[請求項6]
 支持体が、生分解性ポリマーからなるナノファイバーを含有してなる細胞培養担体である、請求項2から5のいずれか一項に記載の細胞シート。

[請求項7]
 腎臓病の治療に用いるための、平均細胞密度が1.0×10 3細胞/cm 2~3.0×10 4細胞/cm 2の間葉系幹細胞をその表面に有する、請求項1から6のいずれか一項に記載の細胞シート。

[請求項8]
 腎臓の線維被膜下に適用するための、請求項7に記載の細胞シート。

[請求項9]
 脳損傷又は神経変性疾患の治療に用いるための、平均密度が0.5×10 3細胞/cm 2~1.5×10 4細胞/cm 2の間葉系幹細胞をその表面に有する、請求項1から6のいずれか一項に記載の細胞シート。

[請求項10]
 脳の損傷部位、変性部位又はそれらの近傍に適用するための、請求項9に記載の細胞シート。

[請求項11]
 間葉系幹細胞が骨髄又は脂肪組織由来の間葉系幹細胞である、請求項1から10のいずれか一項に記載の細胞シート。

[請求項12]
 間葉系幹細胞が疾患を有する対象から分離された間葉系幹細胞である、請求項1から11のいずれか一項に記載の細胞シート。

[請求項13]
 ファイバーからなる3次元構造を有する細胞培養担体上に、間葉系幹細胞を3.0×10 5細胞/cm 2以下の細胞数で播種する工程、及び
 間葉系幹細胞を培養して、平均細胞密度が3.0×10 4細胞/cm 2以下の細胞シートを調製する工程
を含む、生体移植用細胞シートの製造方法。

[請求項14]
 細胞培養担体が、ナノメートル~マイクロメートル単位の平均繊維径を有するファイバーによって細胞との接触面上に形成された開口部を有する、請求項13に記載の製造方法。

[請求項15]
 開口部の平均径が500nm~1000μmである、請求項14に記載の製造方法。

[請求項16]
 細胞培養担体が、生分解性ポリマーからなるナノファイバーを含有してなる細胞培養担体である、請求項13から15のいずれか一項に記載の製造方法。

[請求項17]
 間葉系幹細胞が骨髄又は脂肪組織由来の間葉系幹細胞である、請求項13から16のいずれか一項に記載の製造方法。

[請求項18]
 間葉系幹細胞が疾患を有する対象から分離された間葉系幹細胞である、請求項13から17のいずれか一項に記載の製造方法。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • SAPPORO MEDICAL UNIVERSITY
  • Inventor
  • CHIKENJI Takako
  • FUJIMIYA Mineko
  • SAITO Yuki
  • NAKANO Masako
  • KONARI Naoto
  • OTANI Miho
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 DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW 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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