Cell-releasing agent and method of releasing cell sheet
A cell-sheet releasing agent of the present invention contains an aminated polyrotaxane. The polyrotaxane constituting the skeleton of the cell-releasing agent of the present invention has a structure in which cavities of a plurality of cyclic molecules are threaded onto a linear molecule and both terminals of the linear molecule have a bulky cap bonded thereto so that the cyclic molecules are not dethreaded from the linear molecule. Furthermore, the aminated polyrotaxane contained in the cell-releasing agent of the present invention is a compound in which at least some of hydroxy groups in the cyclodextrin structure contained in the polyrotaxane are each substituted with a substituent having an amino group. According to this cell-sheet releasing agent, cultured cells anchored to the surface of a container can be released without damaging the cells and without controlling the temperature.
1. A method of releasing a cell sheet comprising:
a culture step of anchoring cells to the surface of a container and culturing the cells to form a cell sheet; and
a releasing step of releasing the cell sheet from the surface of the container using a culture medium in which a cell-releasing agent comprising an aminated polyrotaxane is dissolved, after the culture step,
wherein the aminated polyrotaxane is a compound in which at least some of hydroxy groups in the cyclodextrin structure contained in the polyrotaxane are each substituted with a substituent having an amino group.
2. The method according to claim 1 , wherein the polyrotaxane is a compound in which cavities of a plurality of the cyclodextrin are threaded onto a linear molecule and both terminals of the linear molecule are each introduced with a biocompatible group having a bulky substituent with a hydrolyzable bond therebetween.
3. The method according to claim 2 , wherein the substituent having an amino group is represented by —OOCNH-A-N R 1 R 2 wherein A represents a linear or branched hydrocarbon chain, and R 1 and R 2 are the same or different and each represent a hydrogen atom or a linear or branched hydrocarbon group.
4. The method according to claim 3 , wherein A is a linear or branched hydrocarbon chain having 2 to 4 carbon atoms.
5. The method according to claim 3 , wherein R 1 and R 2 are the same or different and each represent a hydrogen atom or a linear or branched hydrocarbon group having 1 to 5 carbon atoms.
6. The method according to claim 1 , wherein the substituent having an amino group is represented by —OOCNH-A-NR 1 R 2 wherein A represents a linear or branched hydrocarbon chain, and R 1 and R 2 are the same or different and each represent a hydrogen atom or a linear or branched hydrocarbon group.
7. The method according to claim 6 , wherein A is a linear or branched hydrocarbon chain having 2 to 4 carbon atoms.
8. The method according to claim 6 , wherein R 1 and R 2 are the same or different and each represent a hydrogen atom or a linear or branched hydrocarbon group having 1 to 5 carbon atoms.
9. The method of releasing a cell sheet according to claim 1 , further comprising:
a recovery step of recovering the released cell sheet.