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Monoclonal antibody K312-based depletion of pluripotent cells from differentiated stem cell progeny prevents teratoma formation
Jongjin Park1 (Postdoctoral researcher), Dong Gwang Lee1 (Research engineer), Na Geum Lee1 (Graduate student), Min-Gi Kwon1,2 (Graduate student), Yeon Sung Son1 (Graduate student), Mi-Young Son3 (Principal researcher), Kwang-Hee Bae4 (Principal researcher), Jangwook Lee1 (Senior researcher), Jong-Gil Park1 (Senior researcher), Nam-Kyung Lee 1 (Associate researcher), Jeong-Ki Min 1,2,* (Principal researcher)
1Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB),
2Department of Biomolecular Science, Korea University of Science and Technology (UST),
3Stem Cell Convergence Research Center and 4Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)
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.
Abstract, Accepted Manuscript(in press) [Submitted on July 9, 2021, Accepted on September 7, 2021]
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