Nongenetic method for purifying stem cell-derived cardiomyocytes.
Several applications of pluripotent stem cell (PSC)-derived cardiomyocytes require elimination of undifferentiated cells. A major limitation for cardiomyocyte purification is the lack of easy and specific cell marking techniques. We found that a fluorescent dye that labels mitochondria, tetramethylrhodamine methyl ester perchlorate, could be used to selectively mark embryonic and neonatal rat cardiomyocytes, as well as mouse, marmoset and human PSC-derived cardiomyocytes, and that the cells could subsequently be enriched (>99% purity) by fluorescence-activated cell sorting. Purified cardiomyocytes transplanted into testes did not induce teratoma formation. Moreover, aggregate formation of PSC-derived cardiomyocytes through homophilic cell-cell adhesion improved their survival in the immunodeficient mouse heart. Our approaches will aid in the future success of using PSC-derived cardiomyocytes for basic and clinical applications.
Hattori
Fumiyuki
F
Department of Regenerative Medicine and Advanced Cardiac Therapeutics, Keio University School of Medicine, Tokyo, Japan.
Chen
Hao
H
Yamashita
Hiromi
H
Tohyama
Shugo
S
Satoh
Yu-Suke
YS
Yuasa
Shinsuke
S
Li
Weizhen
W
Yamakawa
Hiroyuki
H
Tanaka
Tomofumi
T
Onitsuka
Takeshi
T
Shimoji
Kenichiro
K
Ohno
Yohei
Y
Egashira
Toru
T
Kaneda
Ruri
R
Murata
Mitsushige
M
Hidaka
Kyoko
K
Morisaki
Takayuki
T
Sasaki
Erika
E
Suzuki
Takeshi
T
Sano
Motoaki
M
Makino
Shinji
S
Oikawa
Shinzo
S
Fukuda
Keiichi
K
D000818:Animals; D000831:Animals, Newborn; D002144:Callithrix; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D017690:Cell Transplantation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000032:analysis; D006321:Heart / Q000196:embryology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008928:Mitochondria / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D051381:Rats; D012235:Rhodamines / Q000032:analysis; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D012235:Rhodamines; C401833:tetramethylrhodamine methyl ester