US2014106432A1PendingUtilityA1

Cell mass capable of serving as a primitive organ-like structure comprised of a plurality of cell types of somatic origin

Assignee: SHISEIDO CO LTDPriority: Jun 27, 2006Filed: Sep 18, 2013Published: Apr 17, 2014
Est. expiryJun 27, 2026(expired)· nominal 20-yr term from priority
C12N 2501/415C12N 5/0627C12N 5/0666C12N 5/0629C12N 5/0625C12N 5/0697A61L 27/00C12N 2501/40C12N 5/06C12N 5/00
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Claims

Abstract

The present invention provides a method of producing a cell mass capable of serving as a primitive organ-like structure comprised of a plurality of somatic cell types of somatic origin, comprising: preparing cultures containing the plurality of types of somatic cells; mixing the plurality of types of somatic cell cultures followed by adding a Wnt signal activator to the mixed cell culture; subjecting the culture containing the Wnt signal activator to non-plate contact culturing over a predetermined time period; and replacing the medium of the culture cultured by the non-plate contact culturing with medium not containing Wnt signal activator and further culturing for a predetermined time period; wherein, at least one type of the plurality of somatic cells is maintained in an undifferentiated state.

Claims

exact text as granted — not AI-modified
1 - 14 . (canceled) 
     
     
         15 . A method of producing a cell mass capable of serving as a primitive organ-like structure comprised of combinations of outer root sheath (ORS) cells and mesenchymal somatic cells, comprising:
 preparing cultures containing ORS cells and mesenchymal somatic cells;   mixing said ORS cells and mesenchymal somatic cells cultures to obtain a mixed cell culture;   adding a Wnt signal activator to the mixed cell culture, wherein the Wnt signal activator inhibits glycogen synthase-kinase-3 (GSK-3) activity;   culturing the mixed cell culture containing the Wnt signal activator in no-contact culture plates by hanging drop method or spheroid formation method over a predetermined time period; and   replacing the medium of the culture cultured by the hanging drop method or spheroid formation method with medium not containing Wnt signal activator and further culturing for a predetermined time period;   wherein, at least one type of the ORS cells and mesencymal somatic cells is maintained in an undifferentiated state.   
     
     
         16 . The method according to  claim 15 , wherein hair follicles having a hair follicle inducing function are formed from the cell mass. 
     
     
         17 . The method according to  claim 15 , wherein the Wnt signal activator is 6-bromoindirubin-3′-oxime (BIO). 
     
     
         18 . The method according to  claim 15 , wherein the Wnt signal activator demonstrating an inhibitory activity on GSK-3 is selected from bis-indolo(indirubin) compound, (2′Z,3′E)-6-bromoindirubin-3′-oxime, (2′Z,3′E)-6-bromoindirubin-3′-acetoxime, 4-benzyl-2-ethyl-1,2,4-thiadizolidine-3,5-dione, 2,4-dibenzyl-5-oxothiazolidine-3-thione, 2-chloro-1-(4,4-dibromo-thiophen-2-yl)-ethanone, [alpha]-4-dibromoacetophenone, N-(4-methoxybenzyl)-N′-(5-nitro-1,3-thiazol-2-yl)urea) and a GSK-3[beta] peptide inhibitor.

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