کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10903881 | 1086537 | 2015 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Dynamic heterogeneity of DNA methylation and hydroxymethylation in embryonic stem cell populations captured by single-cell 3D high-content analysis
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
DMEMaFGFSOX17FOXA2dSTORMCDH1LIF5hmCgDNACMIIPSCDAPITETHCAForkhead box A2Three-dimensionalFBS4′,6-diamidino-2-phenylindole - 4 '، 6-دیامیدینو-2-فنیلینولOct-4 - 4 اکتبر5mC - 5 سانتی متر5-Methylcytosine - 5-متیل سیتوزین5-hydroxymethylcytosine - 5-هیدروکسی متیل سیتوزینDulbecco׳s modified Eagle׳s medium - Dilbecco's modified Eagle's mediumiGFR - IGFRPCA - PCAPrinciple component analysis - تجزیه و تحلیل اجزای اصلHigh-content analysis - تجزیه و تحلیل با محتوای بالاSuper-resolution imaging - تصویربرداری با وضوح عالیESC - خروجfetal bovine serum - سرم جنین گاوEmbryonic stem cell - سلول های بنیادی جنینیInduced pluripotent stem cell - سلول های بنیادی پلوروپتوژن منجر شده استleukemia inhibitory factor - عامل مهارکننده لوکمیAcidic fibroblast growth factor - فاکتور رشد فیبروبلاست اسیدoctamer-binding transcription factor 4 - فاکتور رونویسی اتصال دهنده octamer 4DNA methylation - متیلاسیون DNAdirect stochastic optical reconstruction microscopy - میکروسکوپ بازسازی اپتیک تصادفی مستقیمhydroxymethylcytosine - هیدروکسی متیل سیتوزینinsulin-like growth factor 1 receptor - گیرنده فاکتور رشد 1 مانند انسولین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
تحقیقات سرطان
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Cell-surface markers and transcription factors are being used in the assessment of stem cell fate and therapeutic safety, but display significant variability in stem cell cultures. We assessed nuclear patterns of 5-hydroxymethylcytosine (5hmC, associated with pluripotency), a second important epigenetic mark, and its combination with 5-methylcytosine (5mC, associated with differentiation), also in comparison to more established markers of pluripotency (Oct-4) and endodermal differentiation (FoxA2, Sox17) in mouse embryonic stem cells (mESC) over a 10-day differentiation course in vitro: by means of confocal and super-resolution imaging together with 3D high-content analysis, an essential tool in single-cell screening. In summary: 1) We did not measure any significant correlation of putative markers with global 5mC or 5hmC. 2) While average Oct-4 levels stagnated on a cell-population base (0.015 lnIU/day), Sox17 and FoxA2 increased 22-fold and 3-fold faster, respectively (Sox17: 0.343 lnIU/day; FoxA2: 0.046 lnIU/day). In comparison, global DNA methylation levels increased 4-fold faster (0.068 lnIU/day), and global hydroxymethylation declined at 0.046 lnIU/day, both with a better explanation of the temporal profile. 3) This progression was concomitant with the occurrence of distinct nuclear codistribution patterns that represented a heterogeneous spectrum of states in differentiation; converging to three major coexisting 5mC/5hmC phenotypes by day 10: 5hmC+/5mCâ, 5hmC+/5mC+, and 5hmCâ/5mC+ cells. 4) Using optical nanoscopy we could delineate the respective topologies of 5mC/5hmC colocalization in subregions of nuclear DNA: in the majority of 5hmC+/5mC+ cells 5hmC and 5mC predominantly occupied mutually exclusive territories resembling euchromatic and heterochromatic regions, respectively. Simultaneously, in a smaller subset of cells we observed a tighter colocalization of the two cytosine variants, presumably delineating chromatin domains in remodeling. We conclude that 1) 5mC emerges as the most differential marker in our model system. 2) However, the combined enrollment of the two DNA modifications provided higher-definition screening and lead to the identification of cell subpopulations based on differential 5hmC/5mC phenotypes corresponding to different 5hmC/5mC ratios. The results encourage: a) assessing the regenerative potential of early-endodermal cells enriched for the three DNA methylation/hydroxymethylation categories, and b) exploring the universality of this type of epigenetic phenotyping across other lineage-specific differentiations.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Experimental Cell Research - Volume 332, Issue 2, 15 March 2015, Pages 190-201
Journal: Experimental Cell Research - Volume 332, Issue 2, 15 March 2015, Pages 190-201
نویسندگان
Jian Tajbakhsh, Darko Stefanovski, George Tang, Kolja Wawrowsky, Naiyou Liu, Jeffrey H. Fair,