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Fig. 1 | Translational Neurodegeneration

Fig. 1

From: Direct conversion of mouse astrocytes into neural progenitor cells and specific lineages of neurons

Fig. 1

Reprogramming of mouse astrocytes into AiNPCs. a Nestin:EGFP transgenic mice were imaged under IVIS optical imaging system. b Schematics for the reprogramming of astrocytes into AiNPCs. c, d Nestin-EGFP+ neurospheres were derived from E15.5 Nestin:EGFP transgenic mouse cortices in suspension culture. e, f Primary astrocyte cultures were positive for GFAP, but negative for Sox2 and Nestin. g-n Representative bright field and fluorescent pictures of cultures during generation of GFP+ NPC-like cells were shown. Step 1, day 0, astrocytes were in adherent culture before retrovirus-mediated Foxg1 + Sox2 + Brn2 transduction (g, h). Step 2, day 28, the cultures formed multiple bulged Nestin-EGFP+ colonies (i, j). Step 3 & 4, Nestin-EGFP+ primary neurospheres appeared and NPC-like cells migrated from neurospheres during adherent culture in poly-D-lysine- and fibronectin-coated dishes (k, l). Mock treated astrocytes showed minimum levels of Nestin-EGFP (m, n). o RNA of astrocytes and AiNPCs was collected and expression of NPC- and glial-specific genes was analyzed using qPCR. Data were normalized to GAPDH and presented as fold change compared with astrocytes. p, q AiNPCs were positive for Ki67 and Sox2. Scale bars represent 10 μm (c-n, p, q). Error bars denote s.d. from triplicate measurements (o)

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