TY - JOUR
T1 - Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media
AU - Tateno, Hiroaki
AU - Hiemori, Keiko
AU - Hirayasu, Kazunari
AU - Sougawa, Nagako
AU - Fukuda, Masakazu
AU - Warashina, Masaki
AU - Amano, Makoto
AU - Funakoshi, Taku
AU - Sadamura, Yoshifusa
AU - Miyagawa, Shigeru
AU - Saito, Atsuhiro
AU - Sawa, Yoshiki
AU - Shofuda, Tomoko
AU - Sumida, Miho
AU - Kanemura, Yonehiro
AU - Nakamura, Masaya
AU - Okano, Hideyuki
AU - Onuma, Yasuko
AU - Ito, Yuzuru
AU - Asashima, Makoto
AU - Hirabayashi, Jun
N1 - Publisher Copyright:
© 2017 The Japanese Society for Regenerative Medicine
PY - 2017/6/1
Y1 - 2017/6/1
N2 - Human pluripotent stem cells are considered to be ideal cell sources for regenerative medicine, but their clinical and industrial application is hindered by their tumorigenic potential. Previously we have identified a pluripotent stem cell-specific lectin rBC2LCN recognizing podocalyxin as a cell surface ligand. More recently, podocalyxin was found to be a soluble ligand of rBC2LCN that is secreted specifically from human pluripotent stem cells into cell culture media. Taking advantage of this phenomenon, we have previously developed a sandwich assay targeting the soluble podocalyxin using rBC2LCN as a capturing probe and another lectin rABA as an overlay probe to detect human pluripotent stem cells residing in cell therapy products derived from human pluripotent stem cells. A drawback to this, however, was that cell culture media containing fetal bovine serum was found to cause a substantial background signal to the sandwich assay. To reduce the background and increase the sensitivity, we screened different overlay probes to detect the soluble podocalyxin. Among them, an anti-keratan sulfate monoclonal antibody called R-10G showed the highest sensitivity and provided a low background signal to fetal bovine serum. The established sandwich assay using rBC2LCN and R-10G was proved to be powerful, which allowed the high-sensitive detection of human induced pluripotent stem cells residing among clinical-grade cardiomyocytes and neural stem cells, both derived from human induced pluripotent stem cells. The developed method has a possibility to be a standard technology to detect human induced pluripotent stem cells resided in various types of cell therapy products.
AB - Human pluripotent stem cells are considered to be ideal cell sources for regenerative medicine, but their clinical and industrial application is hindered by their tumorigenic potential. Previously we have identified a pluripotent stem cell-specific lectin rBC2LCN recognizing podocalyxin as a cell surface ligand. More recently, podocalyxin was found to be a soluble ligand of rBC2LCN that is secreted specifically from human pluripotent stem cells into cell culture media. Taking advantage of this phenomenon, we have previously developed a sandwich assay targeting the soluble podocalyxin using rBC2LCN as a capturing probe and another lectin rABA as an overlay probe to detect human pluripotent stem cells residing in cell therapy products derived from human pluripotent stem cells. A drawback to this, however, was that cell culture media containing fetal bovine serum was found to cause a substantial background signal to the sandwich assay. To reduce the background and increase the sensitivity, we screened different overlay probes to detect the soluble podocalyxin. Among them, an anti-keratan sulfate monoclonal antibody called R-10G showed the highest sensitivity and provided a low background signal to fetal bovine serum. The established sandwich assay using rBC2LCN and R-10G was proved to be powerful, which allowed the high-sensitive detection of human induced pluripotent stem cells residing among clinical-grade cardiomyocytes and neural stem cells, both derived from human induced pluripotent stem cells. The developed method has a possibility to be a standard technology to detect human induced pluripotent stem cells resided in various types of cell therapy products.
KW - Glycan
KW - Lectin
KW - Pluripotent stem cells
KW - Regenerative medicine
KW - Tumorigenicity
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U2 - 10.1016/j.reth.2016.12.002
DO - 10.1016/j.reth.2016.12.002
M3 - Article
AN - SCOPUS:85041060396
VL - 6
SP - 1
EP - 8
JO - Regenerative Therapy
JF - Regenerative Therapy
SN - 2352-3204
ER -