NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations
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 conta...
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| Format: | Article |
| Language: | English |
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Elsevier
2018-05-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506118301119 |
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Article |
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DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
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 |
| spellingShingle |
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 NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations Stem Cell Research |
| author_facet |
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 |
| author_sort |
Shailesh Kumar Gupta |
| title |
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| title_short |
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| title_full |
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| title_fullStr |
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| title_full_unstemmed |
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| title_sort |
nkx6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations |
| publisher |
Elsevier |
| series |
Stem Cell Research |
| issn |
1873-5061 |
| publishDate |
2018-05-01 |
| description |
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 human beta cells. Furthermore, functional characterization of the isolated cells demonstrated that glucose-stimulated insulin secretion is mainly confined to the NKX6.1-positive cells. We expect that the NKX6.1-GFP iPSC lines and the results presented here will contribute to the further refinement of differentiation protocols and characterization of hPSC-derived beta cells and motor neurons for disease modelling and cell replacement therapies. Keywords: Human induced pluripotent stem cells, NKX6.1, Reporter cell line, Directed differentiation, hiPSC-derived beta cells |
| url |
http://www.sciencedirect.com/science/article/pii/S1873506118301119 |
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doaj-388e243f0c3f452f8d7167f3e381a0f12020-11-25T00:29:51ZengElsevierStem Cell Research1873-50612018-05-0129220231NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populationsShailesh Kumar Gupta0Agata Wesolowska-Andersen1Anna K. Ringgaard2Himjyot Jaiswal3Luyan Song4Benoit Hastoy5Camilla Ingvorsen6Amir Taheri-Ghahfarokhi7Björn Magnusson8Marcello Maresca9Rikke R. Jensen10Nicola L. Beer11Johannes J. Fels12Lars G. Grunnet13Melissa K. Thomas14Anna L. Gloyn15Ryan Hicks16Mark I. McCarthy17Mattias Hansson18Christian Honoré19Discovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden; Corresponding authors.Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UKDepartment of Stem Cell Biology, Novo Nordisk A/S, DK-2760 Måløv, DenmarkDiscovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, SwedenLilly Research Laboratories, 46285 Indianapolis, IN, USAOxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UKHistology and Imaging, Novo Nordisk A/S, DK-2760 Måløv, DenmarkDiscovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, SwedenDiscovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, SwedenDiscovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, SwedenDepartment of Stem Cell Biology, Novo Nordisk A/S, DK-2760 Måløv, DenmarkOxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UKResearch Bioanalysis, Novo Nordisk A/S, DK-2760 Måløv, DenmarkDepartment of Stem Cell Biology, Novo Nordisk A/S, DK-2760 Måløv, DenmarkLilly Research Laboratories, 46285 Indianapolis, IN, USAOxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UK; Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK; Oxford NIHR Biomedical Research Centre, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UKDiscovery Biology, Discovery Sciences IMED Biotech Unit, AstraZeneca, Gothenburg, SwedenOxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UK; Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK; Oxford NIHR Biomedical Research Centre, Churchill Hospital, Old Road, Headington, Oxford OX3 7LJ, UKStem Cell Research, Novo Nordisk A/S, DK-2760 Måløv, DenmarkDepartment of Stem Cell Biology, Novo Nordisk A/S, DK-2760 Måløv, Denmark; Corresponding authors.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 human beta cells. Furthermore, functional characterization of the isolated cells demonstrated that glucose-stimulated insulin secretion is mainly confined to the NKX6.1-positive cells. We expect that the NKX6.1-GFP iPSC lines and the results presented here will contribute to the further refinement of differentiation protocols and characterization of hPSC-derived beta cells and motor neurons for disease modelling and cell replacement therapies. Keywords: Human induced pluripotent stem cells, NKX6.1, Reporter cell line, Directed differentiation, hiPSC-derived beta cellshttp://www.sciencedirect.com/science/article/pii/S1873506118301119 |