Single-cell protein expression of hiPSC-derived cardiomyocytes using Single-Cell Westerns.
Single-cell protein expression of hiPSC-derived cardiomyocytes using Single-Cell Westerns.
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DOI：
10.1016/j.yjmcc.2020.09.012
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发表时间：
2020-12
影响因子：
5
通讯作者：
Churko JM
中科院分区：
文献类型：
--
作者：
Jabart E;Molho J;Sin K;Stansfield B;Kazmouz SG;Ventro D;Gardner K;Wu JC;Churko JM
将人类体细胞重编程为人类诱导多能干细胞（hiPSC）的能力使研究人员能够在体外产生具有忠实再现患者特定疾病过程和表型的潜力的细胞类型。hiPSC衍生的心肌细胞（hiPSC-CM）提供了用于药物测试的体外患者和疾病特异性模型以及用于治疗心血管疾病的新型治疗方法的发现的希望。虽然已经证明了将hiPSC分化为心肌细胞的方法，但这些分化群体的异质性和不成熟性限制了它们再现人类疾病和相关靶细胞表型的潜力。这些障碍可以通过全面的单细胞表征来克服，以剖析hiPSC-CM的丰富异质性并研究不同细胞命运的来源。在这项研究中，我们优化并验证了一种新的单细胞Western方法来评估hiPSC-CM中的蛋白表达。为了更好地理解心肌细胞分化产生的不同亚群，并随着时间的推移以单细胞分辨率跟踪群体，我们使用单细胞蛋白质印迹测量和定量心肌细胞亚型特异性蛋白（MLC 2 V和MLC 2A）的表达。通过单细胞蛋白质表达和定量来了解它们的异质性，我们可以改进目前的心肌细胞分化方案，产生更能代表体内衍生心肌细胞的hiPSC-CM用于疾病建模，并将hiPSC-CM用于再生医学目的。单细胞蛋白质印迹为单细胞分辨率的蛋白质表达分析提供了一个强大的平台。
The ability to reprogram human somatic cells into human induced pluripotent stem cells (hiPSCs) has enabled researchers to generate cell types in vitro that have the potential to faithfully recapitulate patient-specific disease processes and phenotypes. hiPSC-derived cardiomyocytes (hiPSC-CMs) offer the promise of in vitro patient- and disease-specific models for drug testing and the discovery of novel therapeutic approaches for treating cardiovascular diseases. While methods to differentiate hiPSCs into cardiomyocytes have been demonstrated, the heterogeneity and immaturity of these differentiated populations have restricted their potential in reproducing human disease and the associated target cell phenotypes. These barriers may be overcome through comprehensive single-cell characterization to dissect the rich heterogeneity of hiPSC-CMs and to study the source of varying cell fates. In this study, we optimized and validated a new Single-Cell Western method to assess protein expression in hiPSC-CMs. To better understand distinct subpopulations generated from cardiomyocyte differentiations and to track populations at single-cell resolution over time, we measured and quantified the expression of cardiomyocyte subtype-specific proteins (MLC2V and MLC2A) using Single-Cell Westerns. By understanding their heterogeneity through single-cell protein expression and quantification, we may improve upon current cardiomyocyte differentiation protocols, generate hiPSC-CMs that are more representative of in vivo derived cardiomyocytes for disease modeling, and utilize hiPSC-CMs for regenerative medicine purposes. Single-Cell Westerns provide a robust platform for protein expression analysis at single-cell resolution.
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影响因子：
16.1
作者：
Feric NT;Radisic M
通讯作者：
Radisic M
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：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya
影响因子：
4.8
作者：
Chen, J;Kubalak, SW;Chien, KR
通讯作者：
Chien, KR