Single-cell RNA sequencing of NGN2-induced neurons from human and chimpanzee pluripotent stem cells
We generated induced excitatory sensory neurons (iNeurons, iNs) from chimpanzee and human stem cells by expressing the transcription factor neurogenin 2 (NGN2). Single cell-RNA sequencing was performed to identify gene expressions differing in dendrite and synapse development during iNs maturation of chimpanzees and humans. Briefly, cultured cells were first dissociated and yield a concentration of 500 to 1000 cells/ml to obtain approximately 3000 cells per lane of a 10X microfluidic chip device. Human and chimpanzee cells were pooled for the 10X experiments and separated computationally afterwards. Single cell gene expression libraries were generated using the 10X Chromium Single Cell 3' v2 Kit following the manufacturer's instructions. Quantification and quality control of libraries was performed using a High Sensitivity DNA chip for Agilent's Bioanalyzer and sequenced on a HiSeq2500 in Rapid sequencing mode.