GSE65423
Transcriptomic characterization of a mixed population of pluripotent stem cell derived cerebellar neuron-like progenitors
We set out to ascertain whether it was possible to use PSCs (pluripotent stem cells) to generate a mix of neuronal cell types resembling those in the cerebellum, most notably Purkinje and granule cells. Several groups have previously demonstrated generation of cerebellar-like neuronal cells from murine ESCs (embryonic stem cells), and more recently the Erceg group achieved similar success using human PSCs. Using principles from these studies we successfully induced expression of mid–hindbrain markers EN1 & GBX2 and later MATH1 (a transcription factor which marks rhombic lip/granule cell progenitors) in a mixed neuronal population derived from hPSCs. Further, we show that it is possible to expand these progenitors to produce spontaneously active neuronal cells with morphological similarities to neuronal cells at specific stages of cerebellar development. Based on morphology, electrophysiology, immunohistochemistry and transcriptomic analysis we demonstrate that these cells express behavior, markers and molecular signatures consistent with distinct epochs of cerebellar development spanning from the late embryonic to early postnatal periods. This study describes an in vitro model that provides insight into the early events that drive the formation of the human cerebellum with support from a high-resolution survey of the trancriptomic landscape, including expression of non-coding RNAs and specific transcript isoforms. The generated datasets and accompanying characterization are a valuable resource for researchers to formulate and test hypotheses regarding the molecular players that are involved in the formation of the cerebellum Overall design: Total RNA-seq profiles of undifferentiated iPS cells and cerebellar neuron-like differentiated cells were generated by deep RNA sequencing, in triplicate, using a HiSeq2500 in Rapid Run mode.