Myosin light chain 2-based selection of human iPSC-derived early ventricular cardiac myocytes.
Myosin light chain 2-based selection of human iPSC-derived early ventricular cardiac myocytes.
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DOI：
10.1016/j.scr.2013.09.003
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发表时间：
2013-11
影响因子：
1.2
通讯作者：
Jalife, Jose
中科院分区：
文献类型：
--
作者：
Bizy, Alexandra;Guerrero-Serna, Guadalupe;Hu, Bin;Ponce-Balbuena, Daniela;Willis, B. Cicero;Zarzoso, Manuel;Ramirez, Rafael J.;Sener, Michelle F.;Mundada, Lakshmi V.;Klos, Matthew;Devaney, Eric J.;Vikstrom, Karen L.;Herron, Todd J.;Jalife, Jose
人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）的应用将通过产生特异性心肌细胞（CM）谱系的能力而得到加强。然而，谱系特异性hiPSC-CM的纯化受到缺乏细胞标记技术的限制。在这里，我们已经开发了一个iPSC-CM标记系统，使用重组腺病毒报告结构与心房或心室特异性肌球蛋白轻链-2（MLC-2）启动子。通过荧光激活细胞分选法纯化MLC-2a和MLC-2 v选择的hiPSC-CM，并分析其生化和电生理表型。我们证明，这两个群体的表型在培养中保持稳定，他们表达预期的肌节蛋白，间隙连接蛋白和室特异性转录因子。与MLC-2a细胞相比，MLC-2 v选择的CM具有更大的动作电位幅度和持续时间。此外，通过免疫荧光，我们表明MLC-2同种型表达可用于富集与早期心房和心室肌细胞谱系一致的hiPSC-CM。然而，只有心室肌球蛋白轻链-2启动子能够纯化高度同质的iPSC-CM群体。使用这种方法，现在可以使用iPSC-CM开发心室特异性疾病模型，而心房特异性iPSC-CM培养物可能需要额外的室特异性标记物。
Applications of human induced pluripotent stemcell 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 ventricularmyocyte 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.
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基因纯化的人诱导多能干细胞衍生的心肌细胞单层的同步电压和钙图谱。
影响因子：
20.1
作者：
Lee P;Klos M;Bollensdorff C;Hou L;Ewart P;Kamp TJ;Zhang J;Bizy A;Guerrero-Serna G;Kohl P;Jalife J;Herron TJ
通讯作者：
Herron TJ
DOI：
10.1083/jcb.111.6.2427
发表时间：
1990-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lyons GE;Schiaffino S;Sassoon D;Barton P;Buckingham M
通讯作者：
Buckingham M
影响因子：
10.8
作者：
Fijnvandraat, AC;van Ginneken, ACG;Deprez, RHL
通讯作者：
Deprez, RHL
DOI：
10.1073/pnas.90.11.5157
发表时间：
1993-06-01
影响因子：
11.1
作者：
OBRIEN, TX;LEE, KJ;CHIEN, KR
通讯作者：
CHIEN, KR
影响因子：
2.7
作者：
Nakagawa, O;Nakagawa, M;Srivastava, D
通讯作者：
Srivastava, D