Method for sorting of pluripotent cells

Patent metadata

Field	Value
Publication number	US-9335323-B2
Application number	US-201314370331-A
Country	US
Kind code	B2
Filing date	Jan 7, 2013
Priority date	Jan 6, 2012
Publication date	May 10, 2016
Grant date	May 10, 2016

Abstract

A method for sorting pluripotent cells using a compound which is eliminated from the pluripotent cells through the MDR1 transporter.

First claim

What is claimed is:

1. A method of sorting non-differentiated pluripotent cells from differentiated somatic cells the method comprising: (i) contacting sample cells with a compound which is eliminated from the pluripotent cells through the cellular MDR1 transporter, wherein the sample cells are pluripotent cells induced to differentiate to somatic cells and, wherein the compound has a structure of the formula (I):

wherein R 1 is selected from the group consisting of the formula (II), (III) and (IV):

wherein R 5 , R 6 , R 7 , R 8 and R 9 are respectively selected from the group consisting of hydrogen, halogen, C1 to C6 alkyl, phenyl, —OR 10 and —SR 10 , wherein R 10 is C1 to C6 alkyl or aryl, wherein R 2 , R 3 and R 4 are respectively selected from the group consisting of hydrogen, C1 to C6 alkyl and —(CH2) bill NR 11 R 12 , and m is an integer of 1 to 12; and R 11 and R 12 are respectively selected from the group consisting of hydrogen, C1 to C6 alkyl and chloroacetyl, or R 2 , and R 3 form the cyclic conformation of the formula (V) together with the nitrogen atom to which R, and R3 are bound:

wherein 1 is an integer of 1 to 6, and X is selected from the group consisting of O, S, NH and NR 13 , wherein R 13 is C1 to C6 alky; and wherein the sample cells which have accumulated the compound are the non-differentiated pluripotent cells (ii) detecting and sorting the cells the non-differentiated pluripotent cells.

2. The method of claim 1 , wherein:
R 2 and R 3 are respectively selected from the group consisting of hydrogen, methyl, ethyl, dimethylaminoethyl and 10-chloroacethylaminodecyl or R 2 and R 3 form the cyclic conformation of 1-piperidinyl;
R 4 is methyl;
R 5 is selected from the group consisting of hydrogen, fluorine, methoxy, phenoxy, methylthio, methyl, propyl and phenyl;
R 6 and R 7 are respectively selected from the group consisting of hydrogen, fluorine, chlorine, methoxy and methyl;
R 8 is fluorine;
R 9 is methoxy; and
X is selected from the group consisting of O, S and methylamino.

3. The method of claim 2 , wherein the compound is selected from the group consisting of the compounds shown in Table 1 below:

TABLE 1

Assignees

• Univ Kyoto

Inventors

• Uesugi Motonari
• Hirata Nao
• Murata Asako
• Chang Young-Tae
• Nakatsuji Norio
• Suemori Hirofumi
• Kawase Eihachiro
• Yamauchi Kaori
• Ueda Kazumitsu
• Fujibayashi Yuto
• Yamanaka Shinya
• Nakagawa Masato

Classifications

• G01N2333/70596

  Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705 · CPC title

• C09B11/28

  Pyronines {; Xanthon, thioxanthon, selenoxanthan, telluroxanthon dyes} · CPC title

• G01N2333/914

  Hydrolases (3) · CPC title

• C09B69/109

  containing other specific dyes · CPC title

• G01N33/5073 Primary

  Stem cells · CPC title