Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media

Hiroaki Tateno, Keiko Hiemori, Kazunari Hirayasu, Nagako Sougawa, Masakazu Fukuda, Masaki Warashina, Makoto Amano, Taku Funakoshi, Yoshifusa Sadamura, Shigeru Miyagawa, Atsuhiro Saito, Yoshiki Sawa, Tomoko Shofuda, Miho Sumida, Yonehiro Kanemura, Masaya Nakamura, Hideyuki Okano, Yasuko Onuma, Yuzuru Ito, Makoto Asashima & 1 others Jun Hirabayashi

Research output: Contribution to journal › Article

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalRegenerative Therapy
Volume6
DOIs
Publication statusPublished - 2017 Jun 1

Fingerprint

Pluripotent Stem Cells
Stem cells
Cell culture
Culture Media
Assays
Cell Culture Techniques
Induced Pluripotent Stem Cells
Cell- and Tissue-Based Therapy
Lectins
Keratan Sulfate
Ligands
Regenerative Medicine
Neural Stem Cells
Serum
Cardiac Myocytes
Monoclonal Antibodies
Technology
Monoclonal antibodies
podocalyxin
Industrial applications

Keywords

  • Glycan
  • Lectin
  • Pluripotent stem cells
  • Regenerative medicine
  • Tumorigenicity

ASJC Scopus subject areas

  • Biomedical Engineering
  • Developmental Biology
  • Biomaterials

Cite this

Tateno, H., Hiemori, K., Hirayasu, K., Sougawa, N., Fukuda, M., Warashina, M., ... Hirabayashi, J. (2017). Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media. Regenerative Therapy, 6, 1-8. https://doi.org/10.1016/j.reth.2016.12.002

Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media. / Tateno, Hiroaki; Hiemori, Keiko; Hirayasu, Kazunari; Sougawa, Nagako; Fukuda, Masakazu; Warashina, Masaki; Amano, Makoto; Funakoshi, Taku; Sadamura, Yoshifusa; Miyagawa, Shigeru; Saito, Atsuhiro; Sawa, Yoshiki; Shofuda, Tomoko; Sumida, Miho; Kanemura, Yonehiro; Nakamura, Masaya; Okano, Hideyuki; Onuma, Yasuko; Ito, Yuzuru; Asashima, Makoto; Hirabayashi, Jun.

In: Regenerative Therapy, Vol. 6, 01.06.2017, p. 1-8.

Research output: Contribution to journal › Article

Tateno, H, Hiemori, K, Hirayasu, K, Sougawa, N, Fukuda, M, Warashina, M, Amano, M, Funakoshi, T, Sadamura, Y, Miyagawa, S, Saito, A, Sawa, Y, Shofuda, T, Sumida, M, Kanemura, Y, Nakamura, M, Okano, H, Onuma, Y, Ito, Y, Asashima, M & Hirabayashi, J 2017, 'Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media', Regenerative Therapy, vol. 6, pp. 1-8. https://doi.org/10.1016/j.reth.2016.12.002
Tateno H, Hiemori K, Hirayasu K, Sougawa N, Fukuda M, Warashina M et al. Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media. Regenerative Therapy. 2017 Jun 1;6:1-8. https://doi.org/10.1016/j.reth.2016.12.002
Tateno, Hiroaki ; Hiemori, Keiko ; Hirayasu, Kazunari ; Sougawa, Nagako ; Fukuda, Masakazu ; Warashina, Masaki ; Amano, Makoto ; Funakoshi, Taku ; Sadamura, Yoshifusa ; Miyagawa, Shigeru ; Saito, Atsuhiro ; Sawa, Yoshiki ; Shofuda, Tomoko ; Sumida, Miho ; Kanemura, Yonehiro ; Nakamura, Masaya ; Okano, Hideyuki ; Onuma, Yasuko ; Ito, Yuzuru ; Asashima, Makoto ; Hirabayashi, Jun. / Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media. In: Regenerative Therapy. 2017 ; Vol. 6. pp. 1-8.
@article{3f51b2e00498455c8cff706e81a6c29b,
title = "Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media",
abstract = "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.",
keywords = "Glycan, Lectin, Pluripotent stem cells, Regenerative medicine, Tumorigenicity",
author = "Hiroaki Tateno and Keiko Hiemori and Kazunari Hirayasu and Nagako Sougawa and Masakazu Fukuda and Masaki Warashina and Makoto Amano and Taku Funakoshi and Yoshifusa Sadamura and Shigeru Miyagawa and Atsuhiro Saito and Yoshiki Sawa and Tomoko Shofuda and Miho Sumida and Yonehiro Kanemura and Masaya Nakamura and Hideyuki Okano and Yasuko Onuma and Yuzuru Ito and Makoto Asashima and Jun Hirabayashi",
year = "2017",
month = "6",
day = "1",
doi = "10.1016/j.reth.2016.12.002",
language = "English",
volume = "6",
pages = "1--8",
journal = "Regenerative Therapy",
issn = "2352-3204",
publisher = "Japanese Society of Regenerative Medicine",

}

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

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

UR - http://www.scopus.com/inward/record.url?scp=85041060396&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041060396&partnerID=8YFLogxK

U2 - 10.1016/j.reth.2016.12.002

DO - 10.1016/j.reth.2016.12.002

M3 - Article

VL - 6

SP - 1

EP - 8

JO - Regenerative Therapy

JF - Regenerative Therapy

SN - 2352-3204

ER -

Access to Document