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Derivation, characterization and differentiation of a new human embryonic stem cell line from a Chinese hatched blastocyst assisted by a non-contact laser system

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Abstract

Currently worldwide attention has focused on the derivation of human embryonic stem cells (hESCs) for future therapeutic medicine. However, the majority of existing hESCs are directly or indirectly exposed to non-human materials during their derivation and/or propagation, which greatly restrict their therapeutic potential. Besides the efforts to improve culture systems, the derivation procedure, especially blastocyst manipulation, needs to be optimized. We adopted a non-contact laser-assisted hatching system in combination with sequential culture process to obtain hatched blastocysts as materials for hESC derivation, and derived a hESC line ZJUhES-1 of a Chinese population without exposure to any non-human materials during blastocyst manipulation. ZJUhES-1 satisfies the criteria of pluripotent hESCs: typically morphological characteristics; the expression of alkaline phosphatase, human telomerase reverse transcriptase and multiple hESC-specific markers including SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, OCT-4, Nanog, Rex-1, Sox-2, UTF-1, Connexins 43 and 45, TERF-1 and TERF-2, Glut-1, BCRP-1/ABCG-2, GDF3, LIN28, FGF4, Thy-1, Cripto1/TDGF1, AC133 as well as SMAD1/2/3/5; extended proliferative capacity; maintenance of a stable male karyotype after long-term cultivation; and robust multiple-lineage developmental potentials both in vivo and in vitro. Moreover, ZJUhES-1 has distinct identity revealed from DNA fingerprinting. Our xeno-free blastocyst manipulation procedure may promote the progression toward clinical-grade hESC derivation.

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Authors and Affiliations

  1. College of Life Sciences, Zhejiang University, 310058, Hangzhou, China

    Rongrong Wu, Zhou Tan, Bin Gu & Ming Zhang

  2. In Vitro Fertilization Center of Women’s Hospital, College of Medicine, Zhejiang University, Hangzhou

    Chenming Xu & Fan Jin

  3. Institute of Genetics and Developmental Biology, the Chinese Academy of Sciences, Beijing

    Liangbiao Chen

  4. Central Hospital of Huzhou, Huzhou, China

    Xing Yao

Authors
  1. Rongrong Wu
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  2. Chenming Xu
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  3. Fan Jin
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  4. Zhou Tan
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  5. Bin Gu
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  6. Liangbiao Chen
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  7. Xing Yao
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  8. Ming Zhang
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Correspondence to Ming Zhang.

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These authors contributed equally to this work.

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Wu, R., Xu, C., Jin, F. et al. Derivation, characterization and differentiation of a new human embryonic stem cell line from a Chinese hatched blastocyst assisted by a non-contact laser system. Hum Cell 23, 89–102 (2010). https://doi.org/10.1111/j.1749-0774.2010.00090.x

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  • DOI: https://doi.org/10.1111/j.1749-0774.2010.00090.x

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