A Knockin Reporter Allows Purification and Characterization of mDA Neurons from Heterogeneous Populations

作者：Xia Ninuo^*; Fang Fang; Zhang Pengbo; Cui Jun; Tep Cullison Chhavy; Hamerley Tim; Lee Hyun Joo; Palmer Theo; Bothner Brian; Lee Jin Hyung; Pera Renee Reijo

来源：Cell Reports, 2017, 18(10): 2533-2546.

DOI：10.1016/j.celrep.2017.02.023

Generation of midbrain dopaminergic (mDA) neurons fromhuman pluripotent stemcells provides a platform for inquiry into basic and translational studies of Parkinson's disease (PD). However, heterogeneity in differentiation in vitro makes it difficult to identify mDA neurons in culture or in vivo following transplantation. Here, we report the generation of a human embryonic stem cell (hESC) line with a tyrosine hydroxylase (TH)-RFP (red fluorescent protein) reporter. We validated that RFP faithfully mimicked TH expression during differentiation. Use of this TH-RFP reporter cell line enabled purification of mDA-like neurons from heterogeneous cultures with subsequent characterization of neuron transcriptional and epigenetic programs (global binding profiles of H3K27ac, H3K4me1, and 5-hydroxymethylcytosine [5hmC]) at four different stages of development. We anticipate that the tools and data described here will contribute to the development of mDA neurons for applications in disease modeling and/or drug screening and cell replacement therapies for PD.

• 出版日期2017-3-7