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EXPANSION CULTURE METHOD FOR NEPHRON PROGENITOR CELLS HAVING NEPHRON-FORMING CAPACITY

外国特許コード F170009024
整理番号 (S2015-1763-N0)
掲載日 2017年3月29日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP070396
国際公開番号 WO 2017010448
国際出願日 平成28年7月11日(2016.7.11)
国際公開日 平成29年1月19日(2017.1.19)
優先権データ
  • 特願2015-139271 (2015.7.11) JP
  • 特願2016-076839 (2016.4.6) JP
発明の名称 (英語) EXPANSION CULTURE METHOD FOR NEPHRON PROGENITOR CELLS HAVING NEPHRON-FORMING CAPACITY
発明の概要(英語) The purpose of the present invention is to provide a method for the expansion culture of nephron progenitor cells while maintaining an undifferentiated state. Another purpose of the present invention is to provide an expansion culture method for cultured nephron progenitor cells to have nephron-forming ability. Yet another purpose of the present invention is to provide an expansion culture method for nephron progenitor cells that makes it possible to achieve an excellent expansion rate. The present invention provides a method for culturing and increasing nephron progenitor cells created from isolated mammalian nephron progenitor cells or iPS cells using a medium containing the following compounds: (i) Wnt agonist; (ii) Notch inhibitor; (iii) Bmp; (iv) leukemia inhibitory factor; (v) Rock inhibitor; and (vi) Fgf.
特許請求の範囲(英語) [claim1]
1. Chemical compound below:
(i) Wnt [agonisuto],
(ii) Fgf,
(iii) Leucocyte obstruction factor (LIF),
(iv) Bmp family chemical compound,
(v) Rock inhibiter, and
(vi) Notch inhibiter
Culturing the [nehuron] fore-quarter cell of the mammal origin which is isolated making use of the nutrient medium which is included, the method of increasing.
[claim2]
2. Furthermore, in the claim 1 to which the nutrient medium includes (vii) Tgf-.alpha. method of statement.
[claim3]
3. The aforementioned nutrient medium, includes the Bmp family chemical compound of the density of 1.0ng/ml-20ng/ml, in claim 1 or 2 method of statement.
[claim4]
4. The aforementioned nutrient medium Wnt [agonisuto] of the density of 0.1 .micro.m-5 .micro.m, the Bmp family chemical compound of the density of 1.0ng/ml-20ng/ml, includes LIF of the density of Fgf, and 1ng/ml-30ng/ml of the density of 10-500ng/ml, either of the claim 1-3 in one method of statement.
[claim5]
5. The aforementioned nutrient medium, includes the Rock inhibiter of the density of 1.0 .micro.m-50 .micro.m, and the Notch inhibiter of the density of 1.0 .micro.m-20 .micro.m, in claim 4 method of statement.
[claim6]
6. The aforementioned nutrient medium, includes Tgf.alpha. of the density of 1ng/ml-500, in claim 5 method of statement.
[claim7]
7. The aforementioned Bmp family chemical compound, is chosen from the group which consists of Bmp2, Bmp4, Bmp7, and GDF11, either of the claim 1-6 in one method of statement.
[claim8]
8. Aforementioned Wnt [agonisuto], is chosen from the GSK-3 inhibiter, either of the claim 1-7 in one method of statement.
[claim9]
9. The description above Fgf, is chosen from the group which consists of Fgf1, Fgf2, Fgf9, and Fgf20, in either of the claim 1-8 method of statement.
[claim10]
10. Aforementioned Wnt [agonisuto] is CHIR99021, the description above Fgf is Fgf9 or Fgf2, the description above Bmp is Bmp7, the aforementioned Rock inhibiter is Y27632, the aforementioned Notch inhibiter is DAPT, either of the claim 1-6 in one method of statement.
[claim11]
11. The aforementioned [nehuron] fore-quarter cell, is the ES cell or the [nehuron] fore-quarter cell of iPS cell origin, either of the claim 1-10 in one method of statement.
[claim12]
12. It features that the aforementioned [nehuron] fore-quarter cell is cultured 5 days or more, either of the claim 1-11 in one method of statement.
[claim13]
13. The aforementioned [nehuron] fore-quarter cell, features that it is cultured to the cell number of 4 times or more by comparison with the cell quantity when starting the culture, either of the claim 1-12 in one method of statement.
[claim14]
14. The aforementioned culture, is done on the iMatrix or [huiburonekuchin] coating plate, either of the above-mentioned claim 1-13 in one method of statement.
[claim15]
15. The aforementioned culture, the U bottom is done low with the cell adsorption plate, either of the claim 1-13 in one method of statement.
[claim16]
16. Either of the claim 1-15 the [nehuron] fore-quarter cell which possesses the [nehuron] formation talent which was cultured to one making use of the method of statement.
[claim17]
17. Either of the claim 1-15 to one the method of drawing up three dimensional kidney structure which possesses the glomerulus and the urine narrow tube making use of the [nehuron] fore-quarter cell which was cultured making use of the method of statement.
[claim18]
18. Chemical compound below:
(i) Wnt [agonisuto],
(ii) Fgf,
(iii) Leucocyte obstruction factor (LIF),
(iv) Bmp family chemical compound,
(v) Rock inhibiter, and
(vi) Notch inhibiter
The nutrient medium in order it includes, to make the [nehuron] fore-quarter cell expand.
[claim19]
19. Furthermore, in the claim 18 which includes (vii) Tgf.alpha. the nutrient medium of statement.
[claim20]
20. The Bmp family chemical compound of the density of 1.0ng/ml-20ng/ml is included, in claim 18 or 19 the nutrient medium of statement.
[claim21]
21. 0.1 Wnt [agonisuto] of the density of .micro.m-5 .micro.m, the Bmp family chemical compound of the density of 1.0ng/ml-20ng/ml, LIF of the density of Fgf, and 1ng/ml-30ng/ml of the density of 10-500ng/ml is included, either of the claim 18-20 in one the nutrient medium of statement.
[claim22]
22. The aforementioned nutrient medium, includes the Rock inhibiter of the density of 1.0 .micro.m-50 .micro.m, and the Notch inhibiter of the density of 1.0 .micro.m-20 .micro.m, in claim 21 the nutrient medium of statement.
[claim23]
23. Tgf.alpha. of the density of 1ng/ml-500 is included, in claim 22 the nutrient medium of statement.
[claim24]
24. Bmp family chemical compound, is chosen from the group which consists of Bmp2, Bmp4, Bmp7, and GDF11, either of the claim 18-23 in one the nutrient medium of statement.
[claim25]
25. Aforementioned Wnt [agonisuto], is the GSK-3 inhibiter, either of the claim 18-24 in one the nutrient medium of statement.
[claim26]
26. The description above Fgf, is chosen from the group which consists of Fgf1, Fgf2, Fgf9, and Fgf20, in either of the claim 18-23 the nutrient medium of statement.
[claim27]
27. Aforementioned Wnt [agonisuto] is CHIR99021, the description above Fgf is Fgf9 or Fgf2, the description above Bmp is Bmp7, the aforementioned Rock inhibiter is Y27632, the aforementioned Notch inhibiter is DAPT, either of the claim 18-26 in one the nutrient medium of statement.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • KUMAMOTO UNIVERSITY
  • 発明者(英語)
  • NISHINAKAMURA RYUICHI
  • TANIGAWA SHUNSUKE
国際特許分類(IPC)
指定国 (WO201710448)
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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