TY - JOUR
T1 - Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches
AU - Miki, Kenji
AU - Endo, Kei
AU - Takahashi, Seiya
AU - Funakoshi, Shunsuke
AU - Takei, Ikue
AU - Katayama, Shota
AU - Toyoda, Taro
AU - Kotaka, Maki
AU - Takaki, Tadashi
AU - Umeda, Masayuki
AU - Okubo, Chikako
AU - Nishikawa, Misato
AU - Oishi, Akiko
AU - Narita, Megumi
AU - Miyashita, Ito
AU - Asano, Kanako
AU - Hayashi, Karin
AU - Osafune, Kenji
AU - Yamanaka, Shinya
AU - Saito, Hirohide
AU - Yoshida, Yoshinori
N1 - Funding Information:
The authors are grateful to Gordon Keller and Mark Gagliardi (University Health Network) for providing valuable advice and support for the cardiomyocyte differentiation method. We thank Nanaka Nishimura and Shunnichi Kashida (Kyoto University) for preparing some of the materials and for critical comments on the project, respectively. We also thank Yasuhiro Yamada, Takuya Yamamoto, Callum Parr, and Peter Karagiannis (Kyoto University) for critical reading of the manuscript. We are grateful to Rie Kato, Sayaka Takeshima, Eri Nishikawa, Ryoko Fujiwara, Kyoko Nakahara, Yukiko Nakagawa, and Yoko Miyake for their administrative support. This research was funded by the Japan Society for the Promotion of Science (JSPS) through the “Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST program)” initiated by the Council for Science and Technology Policy (CSTP) (to S.Y.), the Balzan Prize assigned to S.Y. and H.S., Health and Labour Sciences Research Grants from the Ministry of Health Labour and Welfare (to Y.Y.), grants from the Research Center Network for Realization of Regenerative Medicine (to H.S., Y.Y., and S.Y.), and the iPS Cell Research Fund. A part of this study was supported by JSPS KAKENHI grant numbers 24790277 (to Y.Y.), 25870355 (to K.E.), and 23681042 and 24104002 (to H.S.). S.Y. is a scientific advisor of iPS Academia Japan without salary and K.O. is a member without salary of the scientific advisory boards of iPS Portal, Japan. Kyoto University has filed a patent application broadly relevant to this work. K.E., K.M., S.T., Y.Y., and H.S. are the investigators of record listed on the patent application.
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/6/4
Y1 - 2015/6/4
N2 - Isolation of specific cell types, including pluripotent stem cell (PSC)-derived populations, is frequently accomplished using cell surface antigens expressed by the cells of interest. However, specific antigens for many cell types have not been identified, making their isolation difficult. Here, we describe an efficient method for purifying cells based on endogenous miRNA activity. We designed synthetic mRNAs encoding a fluorescent protein tagged with sequences targeted by miRNAs expressed by the cells of interest. These miRNA switches control their translation levels by sensing miRNA activities. Several miRNA switches (miR-1-, miR-208a-, and miR-499a-5p-switches) efficiently purified cardiomyocytes differentiated from human PSCs, and switches encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as miR-126-, miR-122-5p-, and miR-375-switches purified endothelial cells, hepatocytes, and insulin-producing cells differentiated from hPSCs, respectively. Thus, miRNA switches can purify cell populations for which other isolation strategies are unavailable.
AB - Isolation of specific cell types, including pluripotent stem cell (PSC)-derived populations, is frequently accomplished using cell surface antigens expressed by the cells of interest. However, specific antigens for many cell types have not been identified, making their isolation difficult. Here, we describe an efficient method for purifying cells based on endogenous miRNA activity. We designed synthetic mRNAs encoding a fluorescent protein tagged with sequences targeted by miRNAs expressed by the cells of interest. These miRNA switches control their translation levels by sensing miRNA activities. Several miRNA switches (miR-1-, miR-208a-, and miR-499a-5p-switches) efficiently purified cardiomyocytes differentiated from human PSCs, and switches encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as miR-126-, miR-122-5p-, and miR-375-switches purified endothelial cells, hepatocytes, and insulin-producing cells differentiated from hPSCs, respectively. Thus, miRNA switches can purify cell populations for which other isolation strategies are unavailable.
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U2 - 10.1016/j.stem.2015.04.005
DO - 10.1016/j.stem.2015.04.005
M3 - Article
C2 - 26004781
AN - SCOPUS:84945484140
SN - 1934-5909
VL - 16
SP - 699
EP - 711
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 6
ER -