NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations

Publication date: Available online 23 April 2018 Source:Stem Cell Research Author(s): Shailesh Kumar Gupta, Agata Wesolowska-Andersen, Anna K. Ringgaard, Himjyot Jaiswal, Luyan Song, Benoit Hastoy, Camilla Ingvorsen, Amir Taheri-Ghahfarokhi, Björn Magnusson, Marcello Maresca, Rikke R. Jensen, Nicola L. Beer, Johannes J. Fels, Lars G. Grunnet, Melissa K. Thomas, Anna L. Gloyn, Ryan Hicks, Mark I. McCarthy, Mattias Hansson, Christian Honoré Recent studies have reported significant advances in the differentiation of human pluripotent stem cells to clinically relevant cell types such as the insulin producing beta-like cells and motor neurons. However, many of the current differentiation protocols lead to heterogeneous cell cultures containing cell types other than the targeted cell fate. Genetically modified human pluripotent stem cells reporting the expression of specific genes are of great value for differentiation protocol optimization and for the purification of relevant cell populations from heterogeneous cell cultures. Here we present the generation of human induced pluripotent stem cell (iPSC) lines with a GFP reporter inserted in the endogenous NKX6.1 locus. Characterization of the reporter lines demonstrated faithful GFP labelling of NKX6.1 expression during pancreas and motor neuron differentiation. Cell sorting and gene expression profiling by RNA sequencing revealed that NKX6.1-positive cells from pancreatic differentiations closely resemble hu...
Source: Stem Cell Research - Category: Stem Cells Source Type: research