Journal: Cell stem cell
Article Title: Human pallial MGE-type GABAergic interneuron cell therapy for chronic focal epilepsy
doi: 10.1016/j.stem.2023.08.013
Figure Lengend Snippet: (A,B) Simplified schematics of neuronal subtypes derived from the MGE progenitor domain (FOXG1+ DLX1/2+ NKX2-1+), including GABAergic pallial interneurons (pINs) that migrate to the cortex and hippocampus (HC), as well as multiple lineages that remain in the subpallium. The latter include striatal GABAergic and cholinergic INs, cholinergic projection neurons (PN) of the basal telencephalon, and GABAergic PNs of the globus pallidus. Listed genes expressed in the immature neurons play critical roles in the specification of the indicated neuronal subtypes. (C,D) Representative ICC images of unsorted and sorted lot 1 after cryopreservation and thaw show expression of MGE pIN markers (LHX6, MAFB, MAF, ERBB4), and cholinergic neurons (ISL1). (E) Protein quantification across independent lots (n=10 to 18). (F,G) Quantification of GABA (F) and Acetylcholine (G) neurotransmitters in the culture supernatant from independent pIN lots (n=8), undifferentiated hESCs (n=1) and spinal motor neurons (n=1). (H) Cell viability post-thaw from independent pIN lots (n=16 unsorted/sorted pairs). In E-H, each dot is an average of technical replicates (2-3) from independently manufactured lots. All data are expressed as a mean ± SEM.
Article Snippet: Single cell suspension was incubated with a biotinylated primary antibody against human ERBB4 (BAF1131 R&D Systems), followed by incubation with Anti-Biotin MicroBeads (130-105-637, Miltenyi Biotec), and magnetic sorting was performed on the CliniMACS according to manufacturer instructions (Miltenyi Biotec).
Techniques: Derivative Assay, Expressing