Back to results
US 8,895,302 B2Granted
Directed differentiation of primate pluripotent stem cells into functional basal forebrain cholinergic neurons (BFCNs) and medium spiny gabaergic projection neurons
AssigneeWisconsin Alumni Research Foundation
InventorsSu-Chun Zhang, Yan Liu, Lixiang Ma
Life of this patent
Estimating the term…
Abstract
Methods of efficiently converting primate pluripotent stem cells to GABA neurons or cholinergic neurons, as well as applications thereof, are disclosed.
Claims13
A method of creating a population of GABA projection neurons comprising the steps of
- a) culturing primate pluripotent stem cells under serum-free conditions to obtain Pax6+/Sox1− primitive neural stem cells;
- b) culturing the Pax6+/Sox1− primitive neural stem cells to in the presence of 100-200 ng/ml Sonic Hedgehog (SHH) or 0.5-0.75 μM purmorphamine to obtain a cell population comprising at least 80% Meis2+/Gsh2+/Nkx2.1− GABA progenitors, and
- c) culturing the GABA progenitors in the presence of valproic acid (VPA) and neurotrophic factors selected from the group consisting of brain derived neurotrophic factor (BDNF), glial-derived neurotrophic factors (GDNF), insulin-like growth factor (IGF1), and cyclic AMP (cAMP) to obtain a cell population comprising at least 70% GABK+/DARPP32+/Meis2+ GABA projection neurons.
A method of creating a population of basal forebrain cholinergic neurons (BFCNs), comprising the steps of:
- a) culturing primate pluripotent stem cells under serum-free conditions to obtain Pax6+/Sox1− primate neural stem cells;
- b) culturing the primate neural stem cells in the presence of 500-1000 ng/ml SHH or 1.0-1.25 μM purmorphamine to obtain a cell population comprising at least 90% Nkx2.1+/Lhx8+/Meis2− cholinergic progenitors;
- and
- c) culturing the cholinergic progenitors in the presence of nerve growth factor (NGF) to obtain a cell population comprising at least 30% ChAT+/p75+/Nkx2.1+ BFCNs.
1 more element
Dependencies are inferred from the claim text.