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PRODUCTION METHOD FOR ARTIFICIAL CANCER STEM CELL AND INDUCED DIFFERENTIATION METHOD THEREFOR

Foreign code F130007422
File No. S2011-0743-C0
Posted date Jun 20, 2013
Country WIPO
International application number 2012JP063302
International publication number WO 2012165287
Date of international filing May 24, 2012
Date of international publication Dec 6, 2012
Priority data
  • P2011-118557 (May 27, 2011) JP
Title PRODUCTION METHOD FOR ARTIFICIAL CANCER STEM CELL AND INDUCED DIFFERENTIATION METHOD THEREFOR
Abstract Provided are a cancer stem cell and a production method therefor. The production method for a multipotent cancer stem cell includes a step for introducing Oct3/4, Sox-2, Klf-4, and c-Myc genes into immortalized epithelial cells. A multipotent cancer stem cell produced by this method is provided.
Outline of related art and contending technology BACKGROUND ART
Recent advances in stem cell biology studies, blood cancer and solid tumor cancer stem cells in the presence of strongly suggested. Solid cancer, a small subset of cancer stem cells (side population: also referred to as SP) is present, this population can be used in order to maintain the tumor mass, self-replicating exclusively by undifferentiated cancer stem cells that retain the ability while maintaining produce a number of cancer cells. Cancer stem cell-specific targeting it is possible to control factor, efficiently deplete cancer stem cells from the tumor may be. Such therapeutic agents can be developed, can be to cure cancer. However, such to identify the critical controlling factor, in cancer patient's tumor-derived cancer stem cells also gather, in addition, a portion of the cancer cell lines also gather, the fraction is not uniform, and the identification of cancer stem cell-specific inhibitor search cannot be performed. Therefore, it is possible to create pure 100% as long as the cancer stem cells, it is considered that greatly facilitated the study of the above.
The definition of cancer stem cells self-renewal, (1) pluripotency, (2) are implanted into immunodeficient mice the (3) same phenotype of the original cancer cancer forms are reduced. ' Cancer stem cells' term, ability to self-replicate (malignant tumor phenotype is split into a tumor cell populations in various fields as) defined for convenience as having cancer cells. Many tumors, that reproduce within the body, with tumorigenicity transformed into cancer stem cells derived from single cells. However, origins of cancer stem cells is still not known.
Cancer stem cells, on the basis of cell surface marker expression of various identified, tumor mass or from cancer cell lines can be collected using a cell sorter. However in this method a population of cancer stem cells collected and is a heteroatom, with the features of pure cancer stem cells and only a small part of cells, cancer stem cells in the strict cannot.
In solid tumors (SP) and specific sub-population, the nature of functional cancer stem cells has been achieved. It includes self-replicating and differentiation by raising the ability to form the entire tumor. Solid cancer cell populations in a cell having a specific cell surface marker of cancer stem cells by having known. For example, brain tumors (glioblastoma), prostate cancer, colon cancer, CD133 + cells and CD133 - cells separated from the these cancers of the immune-compromised mice to xenograft experiments, also, in the in vitro ability to self-replicate, growth, have the ability to differentiate from, and CD133 + cells is shown to be cancer stem cells (non-patent document 1-3). In addition, the expression of CD44 in breast cancer is high, a lower expression of the CD24 fractions (CD44 (- / low) CD24 (+) ) have the property of cancer stem cells (non-patent document 4) shown.
However, these cell surface markers of cancer stem cells relies on the separation of, cancer stem cells are separated from the 100% cannot be said to be, in addition, isolated cells in the maintenance or amplification is very difficult. Therefore, it is possible to create artificially cancer stem cells have been developed.
Miyoshi et al. recently, various cancer cells of the gastrointestinal system (cancer cell line) expressing reprogramming factors in cancer stem cells by the induction of successful (non-patent document 5) reports.
However, these cancer cell lines is already been transformed (malignancy, cancer) a number of genes and a mutation or a chromosomal abnormalities has been achieved. Therefore a homogenous cell population which is not configured, drug screening and biomarker considered not suitable for search. Therefore, the cells are defined as pure cancer stem cells is difficult.
On the other hand, human fibroblasts SOX2, OCT4, Klf4, c-Myc yamanaka et al. 4 of the two genes with a retroviral vector can be introduced, (Embryonic stem cells embryonic stem cells: cell ES) induced pluripotent stem cell similar to induce (induced pluripotent stem cell, iPS cell) (non-patent document 6) shown.
However, these 4 human genes introduced will not an immortalized cell, one of these 4 gene introduction, as well as the initialization of the cell, is simultaneously thereby causing cancer, human cancer stem cells are shown it to be formed is not reported.
Scope of claims (In Japanese)請求の範囲 [請求項1]
Oct3/4、Sox-2、Klf-4及びc-Myc遺伝子を不死化上皮細胞に導入することを含む、多能性癌幹細胞の作製方法。

[請求項2]
請求項1記載の方法で作製した多能性癌幹細胞。

[請求項3]
請求項2記載の多能性癌幹細胞を分化させることを含む、癌幹細胞の作製方法。

[請求項4]
請求項3記載の方法で作製した癌幹細胞。

[請求項5]
請求項4記載の癌幹細胞を分化させることを含む、癌細胞の作製方法。

[請求項6]
請求項5記載の方法で作製した癌細胞。

[請求項7]
請求項2、4又は6記載の細胞を用いて、抗癌作用を有する物質をスクリーニングする方法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • Public University Corporation Yokohama City University
  • Inventor
  • RYO, Akihide
  • NISHI, Mayuko
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH 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 IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PE PG PH PL PT QA RO RS RU RW 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 ML MR NE SN TD TG
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