Transcriptional Profiling of Xenogeneic Transplants: Examining Human Pluripotent Stem Cell-Derived Grafts in the Rodent Brain
Christopher R Bye, Vanessa Penna, Isabelle R de Luzy, Carlos W Gantner, Cameron PJ Hunt, Lachlan H Thompson, Clare L Parish
Stem Cell Reports | CELL PRESS | Published : 2019
Human pluripotent stem cells are a valuable resource for transplantation, yet our ability to profile xenografts is largely limited to low-throughput immunohistochemical analysis by difficulties in readily isolating grafts for transcriptomic and/or proteomic profiling. Here, we present a simple methodology utilizing differences in the RNA sequence between species to discriminate xenograft from host gene expression (using qPCR or RNA sequencing [RNA-seq]). To demonstrate the approach, we assessed grafts of undifferentiated human stem cells and neural progenitors in the rodent brain. Xenograft-specific qPCR provided sensitive detection of proliferative cells, and identified germ layer markers a..View full abstract
Awarded by National Health and Medical Research Council Australia
The authors thank Professor Alicia Oshlack, Dr. Jessica Kauhausen, Ms. Mong Tien, and Ms. Brianna Xuerub for their expert technical assistance, and acknowledge the support of the flow cytometry facility at the Melbourne Brain Centre and the MCRI/VCGS Sequencing Service and Development Platform. We thank Vinicius R.M. Carneiro for his assistance in software development to screen transcript homology. V.P. and I.R.d.L. were supported by The University of Melbourne International Scholarships, Australia. C.W.G. is supported by an Australian Postgraduate Award. C.L.P. was supported by a Senior Medical Research Fellowship provided by the Viertel Charitable Foundation, Australia. This research was funded by the National Health and Medical Research Council Australia project grant APP1102704 and Stem Cells Australia. The Florey Institute of Neuroscience and Mental Health acknowledges strong support from the Victorian Government and in particular the funding from the Operational Infrastructure Support Grant.