DEV02541 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Figure 1 - Fig. S1. Culture paradigm for the generation of AHNPs. Progenitors were isolated from a broad range of forebrain locations (green, representative biopsy sites shown for established cell lines) and cultured as mitogen-dependent adherent cultures. Previously described neurosphere-forming cells were isolated from periventricular (red) and hippocampal tissue, and were clonally expanded in nonadhesive culture. Differentiation of neurospheres yields all CNS cell types, while application of ‘neuronization’ conditions induces the formation of neurons (but not oligodendrocytes) in adherent progenitors.

• Supplemental Figure 2 -

  Fig. S2. Distribution and phenotype of transplanyted progenitors. Distribution of SVZ and cortically implanted AHNPs 7 days after transplantation. Both SVZ and cortically implanted AHNPs exhibit modest migration on the anteroposterior axis. While SVZ implanted AHNPs exhibit limited migration from the ventricular surface into the cortex, AHNPs transplanted into the cortex exhibit strong bidirectional migration on the mediolateral axis. Immunophenotyping of AHNPs on the primary engrafting section identifying astrocytes (GFAP⁺), neurons (β-III-tubulin⁺) or undifferentiated cells (GFAP⁻, β-III-tubulin⁻). Data are presented as an average percentage (n=3 animals per experimental condition).

• Supplemental Figure 3 -

  Fig. S3. Optimized lentiviral infection of AHNPs. (A) Seven days post-infection AHNPs were sorted for eGFP expression against untreated cells using standard fluorescence gating. (B) Following FACS analysis, plated cells remain viable and continue to robustly express eGFP. (C) Gated threshold measurements of eGFP expression at 5-20 multiplicities of infection (moi).