Systems and methods for screening for modulators of neural differentiation
OwnerThomas Jessell, Hynek Wichterle +2inventors
InventorsThomas Jessell, Hynek Wichterle, Sara Wilson, Ivo Lieberam
Life of this patent
Estimating the term…
Abstract
The present invention provides in vitro systems for use in identifying modulators of neural differentiation. Also provided are modulators identified by these systems. The present invention further provides methods for identifying a modulator of neural differentiation, a modulator of a Wnt signalling pathway, a modulator of Wnt-dependent neural differentiation, a modulator of a BMP signalling pathway, a modulator of BMP-dependent neural differentiation, a modulator of a Hh signalling pathway, and a modulator of Hh-dependent neural differentiation. Also provided are modulators identified by these methods.
Claims80
An in vitro system for use in identifying a modulator of neural differentiation, comprising:
- (a) a collection of embryonic stem cells;
- (b) a rostralizing or caudalizing embryonic signalling factor;
- and
- (c) optionally, a dorsalizing or ventralizing embryonic signalling factor.
An in vitro system for use in identifying a modulator of neural differentiation, comprising a collection of embryonic stem cells that has been contacted with:
- (a) a rostralizing or caudalizing embryonic signalling factor, to produce a collection of neural progenitor cells;
- and
- (b) optionally, a dorsalizing or ventralizing embryonic signalling factor.
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a collection of embryonic stem cells;
- (b) contacting the embryonic stem cells simultaneously with a rostralizing or caudalizing embryonic signalling factor and a dorsalizing or ventralizing embryonic signalling factor, in the presence of a candidate modulator;
- and
- (c) determining if the candidate modulator modulates neural differentiation of the embryonic stem cells.
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a first collection of embryonic stem cells and a second collection of embryonic stem cells;
- (b) contacting the first collection of embryonic stem cells with a rostralizing or caudalizing embryonic signalling factor and a dorsalizing or ventralizing embryonic signalling factor, in the presence of a candidate modulator;
- (c) contacting the second collection of embryonic stem cells with a rostralizing or caudalizing embryonic signalling factor and a dorsalizing or ventralizing embryonic signalling factor, in the absence of the candidate modulator;
- and
- (d) determining if the candidate modulator in step (b) modulates neural differentiation of embryonic stem cells by comparing neural differentiation of the embryonic stem cells in step (b) with neural differentiation of the embryonic stem cells in step (c).
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a collection of embryonic stem cells;
- (b) contacting the collection of embryonic stem cells with an amount of a rostralizing or caudalizing embryonic signalling factor effective to produce neural progenitor cells;
- (c) contacting the neural progenitor cells with a dorsalizing or ventralizing embryonic signalling factor, in the presence of a candidate modulator;
- and
- (d) determining if the candidate modulator modulates neural differentiation of the embryonic stem cells.
A method for identifying a modulator of a Wnt signalling pathway, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a Wnt signalling pathway;
- (d) contacting the second collection of cells with an activator of a Wnt signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c), wherein modulation of neural differentiation is indicative of modulation of a Wnt signalling pathway.
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a Wnt signalling pathway;
- (d) contacting the second collection of cells with an activator of a Wnt signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c).
A method for identifying a modulator of Wnt-dependent neural differentiation, comprising the steps of:
- (a) obtaining or generating a collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) contacting the collection of cells with a candidate modulator;
- and
- (c) determining if the candidate modulator modulates Wnt-dependent neural differentiation of the cells in the collection.
A method for identifying a modulator of a BMP signalling pathway, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a BMP signalling pathway;
- (d) contacting the second collection of cells with an activator of a BMP signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c), wherein modulation of neural differentiation is indicative of modulation of a BMP signalling pathway.
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a BMP signalling pathway;
- (d) contacting the second collection of cells with an activator of a BMP signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c).
A method for identifying a modulator of BMP-dependent neural differentiation, comprising the steps of:
- (a) obtaining or generating a collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) contacting the collection of cells with a candidate modulator;
- and
- (c) determining if the candidate modulator modulates BMP-dependent neural differentiation of the cells in the collection.
A method for identifying a modulator of a Hh signalling pathway, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a Hh signalling pathway;
- (d) contacting the second collection of cells with an activator of a Hh signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c), wherein modulation of neural differentiation is indicative of modulation of a Hh signalling pathway.
A method for identifying a modulator of neural differentiation, comprising the steps of:
- (a) obtaining or generating a first collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) obtaining or generating a second collection of cells selected from the same group as the cells selected in step (a);
- (c) contacting the first collection of cells with a candidate modulator, in the presence of an activator of a Hh signalling pathway;
- (d) contacting the second collection of cells with an activator of a Hh signalling pathway;
- and
- (e) determining if the candidate modulator in step (b) modulates neural differentiation by comparing neural differentiation of the cells in step (b) with neural differentiation of the cells in step (c).
A method for identifying a modulator of Hh-dependent neural differentiation, comprising the steps of:
- (a) obtaining or generating a collection of cells selected from the group consisting of embryonic stem cells, embryoid bodies, and neural progenitor cells;
- (b) contacting the collection of cells with a candidate modulator;
- and
- (c) determining if the candidate modulator modulates Hh-dependent neural differentiation of the cells in the collection.
Dependencies are inferred from the claim text.