کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8425205 | 1546024 | 2018 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
NKX6.1 induced pluripotent stem cell reporter lines for isolation and analysis of functionally relevant neuronal and pancreas populations
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کلمات کلیدی
PBSNeomycin-resistance geneNK6 homeobox 1Reporter cell lineNkx6.1KRBHGSISPdx1hiPSCHpSCIMINEOHBSSFACSIBMXKClGFPHDR3-isobutyl-1-methylxanthine - 3-ایزوبوتیل-1-متیلکسانتینhESC - hescPancreatic and duodenal homeobox 1 - homeobox پانکراس و دوازدههHomology-directed repair - اصلاح شده با همسانیDirected differentiation - تمایز مستقیمHuman induced pluripotent stem cells - سلول های بنیادی تکامل یافته القا شده توسط انسانhuman embryonic stem cells - سلول های بنیادی جنینی انسانHuman pluripotent stem cells - سلول های بنیادی پلورپوفتون انسانیfluorescence activated cell sorting - فلورسانس سلول فعال فعال سلولPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریHank's balanced salt solution - محلول نمک متعادل هانکGreen fluorescence protein - پروتئین فلورسانس سبزPotassium chloride - کلرید پتاسیمGlucose stimulated insulin secretion - گلوکز ترشح انسولین را تحریک می کند
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
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.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Stem Cell Research - Volume 29, May 2018, Pages 220-231
Journal: Stem Cell Research - Volume 29, May 2018, Pages 220-231
نویسندگان
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,