کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10891299 | 1081998 | 2013 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Myosin light chain 2-based selection of human iPSC-derived early ventricular cardiac myocytes
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کلمات کلیدی
FACSGFPhiPSCAdenovirus - آدنوویروسESC - خروجfluorescence-activated cell sorting - دسته بندی سلول های فعال فلورسنسHuman induced pluripotent stem cell - سلول های بنیادی انعقادی انسانی بشرEmbryonic stem cell - سلول های بنیادی جنینیCardiac myocyte - میوسیت قلبیaction potential - پتانسیل عمل green fluorescent protein - پروتئین فلورسنت سبز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
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چکیده انگلیسی
Applications of human induced pluripotent stem cell derived-cardiac myocytes (hiPSC-CMs) would be strengthened by the ability to generate specific cardiac myocyte (CM) lineages. However, purification of lineage-specific hiPSC-CMs is limited by the lack of cell marking techniques. Here, we have developed an iPSC-CM marking system using recombinant adenoviral reporter constructs with atrial- or ventricular-specific myosin light chain-2 (MLC-2) promoters. MLC-2a and MLC-2v selected hiPSC-CMs were purified by fluorescence-activated cell sorting and their biochemical and electrophysiological phenotypes analyzed. We demonstrate that the phenotype of both populations remained stable in culture and they expressed the expected sarcomeric proteins, gap junction proteins and chamber-specific transcription factors. Compared to MLC-2a cells, MLC-2v selected CMs had larger action potential amplitudes and durations. In addition, by immunofluorescence, we showed that MLC-2 isoform expression can be used to enrich hiPSC-CM consistent with early atrial and ventricular myocyte lineages. However, only the ventricular myosin light chain-2 promoter was able to purify a highly homogeneous population of iPSC-CMs. Using this approach, it is now possible to develop ventricular-specific disease models using iPSC-CMs while atrial-specific iPSC-CM cultures may require additional chamber-specific markers.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Stem Cell Research - Volume 11, Issue 3, November 2013, Pages 1335-1347
Journal: Stem Cell Research - Volume 11, Issue 3, November 2013, Pages 1335-1347
نویسندگان
Alexandra Bizy, Guadalupe Guerrero-Serna, Bin Hu, Daniela Ponce-Balbuena, B. Cicero Willis, Manuel Zarzoso, Rafael J. Ramirez, Michelle F. Sener, Lakshmi V. Mundada, Matthew Klos, Eric J. Devaney, Karen L. Vikstrom, Todd J. Herron, José Jalife,