CELLULES SOUCHES DE BORDURE DE PLAQUE NEURALE DE MAMMIFÈRES CAPABLES DE FORMER DES LIGNÉES DE CELLULES DU TUBE NEURAL ET DE LA CRÊTE NEURALE INCLUANT DES NEURONES CENTRAUX ET PÉRIPHÉRIQUES
Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V.
发明人:
申请号：
EP13700672.2
公开号：
EP2802332B1
申请日:
2013.01.11
申请国别(地区):
EP
年份:
2018
代理人:
摘要：
The present invention relates to a method for producing mammalian neural plate border stem cells (NPBSCs), comprising: (a) differentiation of mammalian pluripotent stem cells by (a-i) culturing mammalian pluripotent stem cells in pluripotent stem cell medium for about 24 to about 96 hours, wherein the pluripotent stem cell medium comprises: (i) an inhibitor of the activin/TGF-² signalling pathway； (ii) an inhibitor of the BMP signalling pathway； (iii) an activator of the canonical WNT signalling pathway； and (iv) an activator of the Hedgehog signalling pathway； subsequently (a-ii) culturing the cells obtained in step (a-i) for about 24 to about 96 hours in a neural medium, wherein the neural medium comprises: (i) an inhibitor of the Activin/TGF-² signalling pathway； (ii) an inhibitor of the BMP signalling pathway； (iii) an activator of the canonical WNT signalling pathway； and (iv) an activator of the Hedgehog signalling pathway； subsequently (a-iii) culturing the cells obtained in step (a-ii) for about 24 to about 96 hours in a neural medium, wherein the neural medium comprises: (i) an activator of the canonical WNT signalling pathway； (ii) an activator of the Hedgehog signalling pathway； and (iii) an inhibitor of oxidation； and (b) plating the obtained differentiated mammalian pluripotent stem cells in NPBSCs expansion medium, wherein the NPBSCs expansion medium comprises (i) an activator of the canonical WNT signalling pathway； (ii) an activator of the Hedgehog signalling pathway； and (iii) an inhibitor of oxidation； and expanding the cells in the NPBSCs expansion medium for about 24 to about 96 hours； (c) splitting the cells obtained in (b) and further expanding the cells in the NPBSCs expansion medium； and (d) repeating step (c) at least two times. The present invention further relates to neural plate border stem cells obtainable by the method of the invention and the use of the cells of the invention in medicine.
