Journal article
Generation of Adrenal Chromaffin-like Cells from Human Pluripotent Stem Cells
KD Abu-Bonsrah, D Zhang, AR Bjorksten, M Dottori, DF Newgreen
Stem Cell Reports | CELL PRESS | Published : 2018
Abstract
Adrenomedullary chromaffin cells are catecholamine (CA)-producing cells originating from trunk neural crest (NC) via sympathoadrenal progenitors (SAPs). We generated NC and SAPs from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in vitro via BMP2/FGF2 exposure, ascertained by qPCR and immunoexpression of SOX10, ASCL1, TFAP2α and PHOX2B, and by fluorescence-activated cell sorting selection for p75NTR and GD2, and confirmed their trunk-like HOX gene expression. We showed that continuing BMP4 and curtailing FGF2 in vitro, augmented with corticosteroid mimetic, induced these cells to upregulate the chromaffin cell-specific marker PNMT and other CA synthesis and sto..
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Awarded by Marshfield Clinic Research Institute
Funding Acknowledgements
We thank Matthew Burton and Paul Lau (Flow Cytometry-MCRI) for their technical advice and support. The alpha TH, B2B1, TFAP2 alpha, and SA1 antibodies were developed and supplied by the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. We would like to also thank Dr. Marthe Howard (University of Toledo) for her advice on antibody selection, A/Prof Alice Pebay for providing the 007 iPSC line, Rachael Chatterton, Centre for Neural Engineering, The University of Melbourne, for her help in culturing the 007 iPSC cell line, Ana Antonic-Baker for her advice on the statistical methods, and Dr. David Elliot, Murdoch Children's Research Institute, for the pSMAD antibody. This research was supported by NHMRC grant 1069757, grant 1050692, Stem Cells Australia (SCA) 2013F-9, Australian Research Council Future Fellowship, The University of Melbourne, and MCRI. Vagal NC-related constructs were contributed to by Foundation for Children 2014-211. We acknowledge the Victorian Government's Operational Infrastructure Support Program to MCRI. K.D.A.-B. holds an IPRS and APA (Int) PhD Scholarship through the Department of Paediatrics, University of Melbourne.