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Figure 1 | Molecular Brain

Figure 1

From: Reprogramming non-human primate somatic cells into functional neuronal cells by defined factors

Figure 1

Schematic overview of the experimental procedure for generation of common marmoset induced neuronal cells from common marmoset embryonic skin fibroblasts. (A) Common marmoset embryonic skin fibroblasts (cjFs) were infected with drug-inducible lentiviral vectors coding a set of induced neuronal (iN) cell factors: ASCL1, BRN2, MYT1L, and NEUROD1. Synapsin reporter lentivirus expressing green fluorescent protein (GFP) or DsRed under the human synapsin I promoter was used to monitor neuronal induction. The lentiviral vector that stably drives rtTA expression under the EF1α promoter was transduced to induce the expression of iN-factors. (B) Time course for common marmoset induced neuronal (cjiN) cell induction. Lentivirus infection was conducted at 0 day in vitro (div) in fibroblast medium containing 10% fetal bovine serum, and cells were exposed to doxycycline (dox) at 1 div for cjiN induction. The culture medium was then replaced at 3 div with dox-containing neural meium composed of N2B27 medium, neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF). For calcium imaging, the cAMP analog, 8-(4-Chlorophenylthio) adenosine 3′, 5′-cyclic monophosphate (8-CPT; 100 μM) [29], was added to promote neuronal maturation and survival.

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