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Title: High-efficiency derivation of human embryonic stem cell lines using a culture system with minimized trophoblast cell proliferation
Authors: Chuti Laowtammathron
Pimjai Chingsuwanrote
Roungsin Choavaratana
Suphadtra Phornwilardsiri
Ketsara Sitthirit
Chidchanok Kaewjunun
Orawan Makemaharn
Papussorn Terbto
Supaporn Waeteekul
Chanchao Lorthongpanich
Yaowalak U-Pratya
Pimonwan Srisook
Pakpoom Kheolamai
Surapol Issaragrisil
Faculty of Medicine, Thammasat University
Faculty of Medicine, Siriraj Hospital, Mahidol University
Keywords: Biochemistry, Genetics and Molecular Biology;Medicine
Issue Date: 11-May-2018
Citation: Stem Cell Research and Therapy. Vol.9, No.1 (2018)
Abstract: © 2018 The Author(s). Background: Due to their extensive self-renewal and multilineage differentiation capacity, human embryonic stem cells (hESCs) have great potential for studying developmental biology, disease modeling, and developing cell replacement therapy. The first hESC line was generated in 1998 by culturing inner cell mass (ICM) cells isolated from human blastocysts using an immunosurgery technique. Since then, many techniques including mechanical ICM isolation, laser dissection, and whole embryo culture have been used to derive hESC lines. However, the hESC derivation efficiency remains low, usually less than 50%, and it requires a large number of human embryos to derive a significant number of hESC lines. Due to a shortage of and restricted access to human embryos, a novel approach with better hESC derivation efficiency is badly needed to decrease the number of embryos used. Methods: We hypothesized that the low hESC derivation efficiency might be due to extensive proliferation of trophoblast (TE) cells which could interfere with ICM proliferation. We therefore developed a methodology to minimize TE cell proliferation by culturing ICM in a feeder-free system for 3 days before transferring them onto feeder cells. Results: This minimized trophoblast cell proliferation (MTP) technique could be successfully used to derive hESCs from normal, abnormal, and frozen-thawed embryos with better derivation efficiency of more than 50% (range 50-100%; median 70%). Conclusions: We successfully developed a better hESC derivation methodology using the "MTP" culture system. This methodology can be effectively used to derive hESCs from both normal and abnormal embryos under feeder-free conditions with higher efficiency when compared with other methodologies. With this methodology, large-scale production of clinical-grade hESCs is feasible.
ISSN: 17576512
Appears in Collections:Scopus 2018

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