Label-free identification and characterization of human pluripotent stem cell-derived cardiomyocytes using second harmonic generation (SHG) microscopy

被引:20
作者
Awasthi, Samir [2 ,5 ]
Matthews, Dennis L. [2 ,3 ]
Li, Ronald A. [4 ,7 ]
Chiamvimonvat, Nipavan [1 ]
Lieu, Deborah K. [1 ]
Chan, James W. [2 ,6 ]
机构
[1] Univ Calif Davis, Dept Internal Med, Davis, CA 95616 USA
[2] Univ Calif Davis, NSF Ctr Biophoton Sci & Technol, Sacramento, CA 95817 USA
[3] Lawrence Livermore Natl Lab, Livermore, CA 94551 USA
[4] Mt Sinai Sch Med, Cardiovasc Res Ctr, New York, NY 10029 USA
[5] Univ Calif Davis, Dept Biomed Engn, Davis, CA 95616 USA
[6] Univ Calif Davis, Dept Pathol & Lab Med, Sacramento, CA 95817 USA
[7] Univ Hong Kong, Stem Cell & Regenerat Med Consortium, Dept Physiol & Med, LKS Fac Med, Hong Kong, Hong Kong, Peoples R China
基金
美国国家科学基金会;
关键词
cardiomyocytes; pluripotent stem cells; second harmonic generation; cell separation; myosin; HEART-CELLS; DIFFERENTIATION; SPECTROSCOPY; ENRICHMENT; THERAPY; CULTURE; TISSUES; MYOSIN;
D O I
10.1002/jbio.201100077
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) are a potentially unlimited source of cardiomyocytes (CMs) for cardiac transplantation therapies. The establishment of pure PSC-CM populations is important for this application, but is hampered by a lack of CM-specific surface markers suitable for their identification and sorting. Contemporary purification techniques are either non-specific or require genetic modification. We report a second harmonic generation (SHG) signal detectable in PSC-CMs that is attributable to sarcomeric myosin, dependent on PSC-CM maturity, and retained while PSC-CMs are in suspension. Our study demonstrates the feasibility of developing a SHG-activated flow cytometer for the non-invasive purification of PSC-CMs. (C) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
引用
收藏
页码:57 / 66
页数:10
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