Specification of functional cranial placode derivatives from human pluripotent stem cells
First Claim
1. A method for inducing differentiation of cells comprisinga) contacting a plurality of starting cells with an inhibitor of Small Mothers Against Decapentaplegic (SMAD) protein signaling (“
- the SMAD inhibitor”
), wherein the starting cells are selected from the group consisting of multipotent cells, pluripotent cells, and a combination thereof; and
b) contacting the cells with a bone morphogenetic protein (BMP);
c) contacting the cells with a compound selected from the group consisting of BRL-54443, parthenolide, phenanthroline, and combinations thereof; and
wherein the cells are contacted with the SMAD inhibitor and the BMP in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells.
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Abstract
Cranial placodes are embryonic structures essential for sensory and endocrine organ development. The efficient derivation of cranial placodes from human pluripotent stem cells is disclosed where the timed removal of the BMP inhibitor Noggin, a component of the dual-SMAD inhibition strategy of neural induction, triggers placode induction at the expense of CNS fates. Further fate specification at the pre-placode stage enables the selective generation of placode-derived trigeminal ganglia capable of in vivo engraftment, mature lens fibers and anterior pituitary hormone-producing cells that upon transplantation produce hormones including, but not limited to, human growth hormone and adrenocortiocotropic hormone in vivo. Alternatively, anterior pituitary hormone-producing cells are generated in cell culture systems in vitro.
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Citations
7 Claims
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1. A method for inducing differentiation of cells comprising
a) contacting a plurality of starting cells with an inhibitor of Small Mothers Against Decapentaplegic (SMAD) protein signaling (“ - the SMAD inhibitor”
), wherein the starting cells are selected from the group consisting of multipotent cells, pluripotent cells, and a combination thereof; andb) contacting the cells with a bone morphogenetic protein (BMP); c) contacting the cells with a compound selected from the group consisting of BRL-54443, parthenolide, phenanthroline, and combinations thereof; and wherein the cells are contacted with the SMAD inhibitor and the BMP in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells. - View Dependent Claims (2, 3, 4, 5, 6)
- the SMAD inhibitor”
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7. A method for inducing differentiation of cells comprising
a) contacting a plurality of starting cells with a first inhibitor of SMAD protein signaling (“ - the first SMAD inhibitor”
), wherein the starting cells are selected from the group consisting of multipotent cells, pluripotent cells, and a combination thereof;b) contacting the cells with a second inhibitor of SMAD protein signaling (“
the second SMAD inhibitor”
); andc) contacting the cells with a compound selected from the group consisting of BRL-54443, parthenolide, phenanthroline, and combinations thereof, wherein the cells are contacted with the first SMAD inhibitor, the second SMAD inhibitor, and the compound in an amount effective to induce detectable expression of SIX1 and PAX6 in the plurality of cells.
- the first SMAD inhibitor”
Specification