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: Key resources table

Article Snippet: WNT pathway inhibition conferred forebrain identity (XAV939 10μM; 3748 Tocris) 105 , 106 and SHH pathway activation induced ventral forebrain MGE-like progenitors (SAG 100nM; 4366 Tocris) 37 , 38 , 40 , 43 , 105 . . During the second MGE progenitor expansion and pallial interneuron commitment phase, a MEK pathway inhibitor was applied to further improve the efficiency, kinetics, and specificity of a pallial-type GABAergic interneuron lineage (PD0325901 2μM; 4192 Tocris).

Techniques: Control, Plasmid Preparation, Recombinant, Software

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: WNT pathway inhibition conferred forebrain identity (XAV939 10μM; 3748 Tocris) 105 , 106 and SHH pathway activation induced ventral forebrain MGE-like progenitors (SAG 100nM; 4366 Tocris) 37 , 38 , 40 , 43 , 105 . . During the second MGE progenitor expansion and pallial interneuron commitment phase, a MEK pathway inhibitor was applied to further improve the efficiency, kinetics, and specificity of a pallial-type GABAergic interneuron lineage (PD0325901 2μM; 4192 Tocris).

Techniques: Derivative Assay, Expressing

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) Expression of immature neuronal marker doublecortin (DCX) (A) and MGE IN marker SST (B) in human cells (HNA+) at 1, 4 and 8.5 MPT. (C) Human-specific axonal marker hTAU showing transplanted cell processes throughout the epileptic HC (a-d), counter-stained with NEUN. (D) Grafted human cells were analyzed by FISH using probes against a housekeeping gene for human-specific glyceraldehyde-3-phosphate dehydrogenase (hGADPH) and the GABAergic marker glutamate decarboxylase 1 (GAD1). Representative image shows cell distribution in the rostral HC with a higher magnification example of the hilus (1). (E) Representative IHC images showing co-labeling of human cells (HNA+, red) with on-target MGE IN markers in green: LHX6, SST, NPY, PV as well as perineuronal nets marker WFA (white). Arrowheads point to specific examples of human cells expressing the markers of interest. Off-target populations, including markers for non-MGE lineage INs (CR), oligodendrocytes (OLIG2), human astrocytes (hGFAP), and proliferating cells (KI67) are shown. (F,G) Quantification of human cell persistence (F) and percent of human cells expressing GAD1 mRNA, and LHX6, SST, KI67, OLIG2 and GFAP proteins (G).

Article Snippet: WNT pathway inhibition conferred forebrain identity (XAV939 10μM; 3748 Tocris) 105 , 106 and SHH pathway activation induced ventral forebrain MGE-like progenitors (SAG 100nM; 4366 Tocris) 37 , 38 , 40 , 43 , 105 . . During the second MGE progenitor expansion and pallial interneuron commitment phase, a MEK pathway inhibitor was applied to further improve the efficiency, kinetics, and specificity of a pallial-type GABAergic interneuron lineage (PD0325901 2μM; 4192 Tocris).

Techniques: Expressing, Marker, Staining, Labeling

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: Samples that were sequenced included day 0 hESCs (n=1), day 14 MGE progenitors (n=1), and week 6 EOP cells (n=3 paired unsorted/sorted lots). (A,B) UMAP (Uniform Manifold Approximation and Projection) visualization of cell clusters in all the samples combined (A), and in each of the separate samples listed (B). (C) Quantification of sample composition by cluster. (D) Feature plots of gene expression. All cells are displayed in light gray, cells with detectable expression are displayed in purple, with darker shade corresponding to higher expression level. (E) Dot plot for key genes that define different cell categories, including general markers of neurons, GABAergic and GE neurons, MGE, MGE pINs, MGE subpallial neurons, POA, CGE, LGE, neuronal progenitors (including MGE progenitor marker NKX2-1), cell cycle, pluripotent, as well as genes associated with glial cells, glutamatergic neurons (Glu), dopaminergic neurons (DA), serotonergic neurons (5HT) and cholinergic neurons (Ach). (F,G) Comparison of in vitro-derived Day 14 progenitors and EOP INs with human developing GE (GW9-18), using the Shi et al.dataset78 as a reference. (F) Prediction scores between 0 and 1 are projected onto day 14 and EOP UMAP. (G) Heatmap showing percentage of cells in each in vitro cluster that are assigned to different GE categories based on prediction scores.

Article Snippet: WNT pathway inhibition conferred forebrain identity (XAV939 10μM; 3748 Tocris) 105 , 106 and SHH pathway activation induced ventral forebrain MGE-like progenitors (SAG 100nM; 4366 Tocris) 37 , 38 , 40 , 43 , 105 . . During the second MGE progenitor expansion and pallial interneuron commitment phase, a MEK pathway inhibitor was applied to further improve the efficiency, kinetics, and specificity of a pallial-type GABAergic interneuron lineage (PD0325901 2μM; 4192 Tocris).

Techniques: Gene Expression, Expressing, Marker, Comparison, In Vitro, Derivative Assay

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: Key resources table

Article Snippet: WNT pathway inhibition conferred forebrain identity (XAV939 10μM; 3748 Tocris) 105 , 106 and SHH pathway activation induced ventral forebrain MGE-like progenitors (SAG 100nM; 4366 Tocris) 37 , 38 , 40 , 43 , 105 . . During the second MGE progenitor expansion and pallial interneuron commitment phase, a MEK pathway inhibitor was applied to further improve the efficiency, kinetics, and specificity of a pallial-type GABAergic interneuron lineage (PD0325901 2μM; 4192 Tocris).

Techniques: Control, Plasmid Preparation, Recombinant, Software

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Induced Resistance to Ofatumumab Mediated Cell Clearance Mechanisms, Including Complement Dependent Cytotoxicity, in Chronic Lymphocytic Leukemia

doi: 10.4049/jimmunol.1302954

Figure Lengend Snippet: CLL cells obtained from patients before treatment (0hr), and then at 8hr and 24hr after the start of the administration of the first dose of ofatumumab (OFA) were tested for in vitro CDC using saturating doses (10 μg/ml) of rituximab (RTX), OFA, or alemtuzumab (ALM)and 10% normal human serum (NHS) as a source of complement. RTX induced low levels of CDC. Compared to OFA CDC in pre-treatment specimens (median 37% cytotoxicity, range 0 – 83) there was a significant decrease in OFA CDC at 8hr (median 0%, range 0 – 5)(p<0.0001) and 24hr (median 0%, range 0 – 32) (p<0.0001). In contrast, ALM CDC was not decreased by in vivo exposure of CLL cells to OFA.

Article Snippet: OFA standards were prepared by serial dilutions of a commercial sample of OFA (GlaxoSmithKline, Research Triangle Park, NC) into pooled normal human serum.

Techniques: In Vitro, In Vivo