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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Monoclonal antibody K312-based depletion of pluripotent cells from differentiated stem cell progeny prevents teratoma formation

  • Park, Jongjin (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Dong Gwang (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Na Geum (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kwon, Min-Gi (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Son, Yeon Sung (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Son, Mi-Young (Stem Cell Convergence Research Center, KRIBB) ;
  • Bae, Kwang-Hee (Metabolic Regulation Research Center, KRIBB) ;
  • Lee, Jangwook (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park, Jong-Gil (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Nam-Kyung (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Min, Jeong-Ki (Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Received : 2021.07.09
  • Accepted : 2021.09.07
  • Published : 2022.03.31
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Abstract

Human pluripotent stem cells (PSCs) have been utilized as a promising source in regenerative medicine. However, the risk of teratoma formation that comes with residual undifferentiated PSCs in differentiated cell populations is most concerning in the clinical use of PSC derivatives. Here, we report that a monoclonal antibody (mAb) targeting PSCs could distinguish undifferentiated PSCs, with potential teratoma-forming activity, from differentiated PSC progeny. A panel of hybridomas generated from mouse immunization with H9 human embryonic stem cells (hESCs) was screened for ESC-specific binding using flow cytometry. A novel mAb, K312, was selected considering its high stem cell-binding activity, and this mAb could bind to several human induced pluripotent stem cells and PSC lines. Cell-binding activity of K312 was markedly decreased as hESCs were differentiated into embryoid bodies or by retinoic acid treatment. In addition, a cell population negatively isolated from undifferentiated or differentiated H9 hESCs via K312 targeting showed a significantly reduced expression of pluripotency markers, including Oct4 and Nanog. Furthermore, K312-based depletion of pluripotent cells from differentiated PSC progeny completely prevented teratoma formation. Therefore, our findings suggest that K312 is utilizable in improving stem cell transplantation safety by specifically distinguishing residual undifferentiated PSCs.

Keywords

Acknowledgement

This research was supported by the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program (KGM5272221) and by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2021R1I1A2057698).

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