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Selective Depletion of SSEA-3- and TRA-1-60-Positive Undifferentiated Human Embryonic Stem Cells by Magnetic Activated Cell Sorter (MACS)

Authors
 Hyun Ok Kim  ;  Yong Joon Huh  ;  Jiho Jang  ;  Youjung Choi  ;  Dong-Wook Kim  ;  Han-Soo Kim 
Citation
 TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Vol.8(2) : 253-261, 2011 
Journal Title
TISSUE ENGINEERING AND REGENERATIVE MEDICINE(조직공학과 재생의학)
ISSN
 1738-2696 
Issue Date
2011
Keywords
MACS ; selective ablation ; monoclonal antibody ; embryonic stem cells ; teratoma
Abstract
The capacity of indefinite self-renewal and pluripotency make human embryonic stem cells (hESCs) and
induced pluripotent stem cells (iPSCs) an attractive source for potential regenerative medicine. However, teratoma
formation is one of the major obstacles for implementing hESC-based therapeutics in a clinical setting. While this
tumorigenic capacity is known to be lost upon lineage differentiation in vitro, there is a potential risk that any residual
undifferentiated hESCs or progenitor cells may form tumors upon transplantation. To ensure the safety of hESCs in
clinical application, we thus explored the magnetic activated cell sorter (MACS) as a tool for separating undifferentiated
hESCs from a mixed population of hESCs and human blood mononuclear cells. Labeled with single or combinations
of two fluorsecein-labeled monoclonal antibodies (SSEA-3 and TRA-1-60) and subsequently stained with
anti-FITC and/or anti-PE magnetic microbeads, cells were subjected to MACS separation. While a reduction of
hESCs by depletion with a single marker was observed, there was still a significant fraction of residual hESCs in the
flow-through fraction. However, two consecutive MACS separations upon simultaneous staining of two different
hESC markers, SSEA-3 and TRA-1-60, completely depleted hESCs, as validated by flow cytometer, real-time PCR
and immunofluoresence microscopic analyses. The maximum efficacy of hESC removal using this protocol was
higher than 99.9%. No teratoma formation was observed in hESC-depleted cell injection to NOD/SCID mice. These
results indicate that the current MACS protocol with two antibodies can efficiently eliminate undifferentiated cells
from differentiated cells and greatly alleviate concerns about tumor formation by hESC-derived cellular therapeutics
Files in This Item:
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Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Physiology (생리학교실) > 1. Journal Papers
Yonsei Authors
Kim, Dong Wook(김동욱) ORCID logo https://orcid.org/0000-0002-5025-1532
Kim, Han Soo(김한수)
Kim, Hyun Ok(김현옥) ORCID logo https://orcid.org/0000-0002-4964-1963
Jang, Ji Ho(장지호) ORCID logo https://orcid.org/0000-0001-5551-3514
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/92763
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