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Hemato-endothelial differentiation from lentiviral-transduced human embryonic stem cells retains durable reporter gene expression under the control of ubiquitin promoter

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Abstract

Human embryonic stem (hES) cells are able to give rise to a variety of cell lineages under specific culture condition. An effective strategy for stable genetic modification in hES cells may provide a powerful tool for study of human embryogenesis and cell-based therapies. However, gene silences are documented in hES cells. In current study, we investigated whether genes controlled under ubiquitin promoter are expressed during hematopoietic-endothelial differentiation in hES cells. Undifferentiated hES cells (H1) were transduced by lentivirus encoding green fluorescent protein (GFP) gene under ubiquitin promoter. GFP-expressing hES cells (GFP-H1) were established after several rounds of mechanical selection under fluorescence microscope. GFP gene was stably expressed in hES cells throughout prolonged (> 50 passages) cultivation, and in differentiated embryo body (EB) and teratoma. Hematopoietic and endothelial markers, including KDR (VEGFR2), CD34, CD31, Tie-2, GATA-1 and GATA-2, were expressed at similar levels during hES cell differentiation in parent hES cells and GFP-H1 hES cells. CD34+ cells isolated from GFP-H1 hES cells were capable to generate hematopoietic colony-forming cells and tubular structure-forming cells. Differentiated GFP-EB formed vasculature structures in a semi-solid sprouting EB model. These results indicated that a transgene under ubiquitin promoter in lentiviral transduced hES cells retained its expression in undifferentiated hES cells and in hES-derived hematopoietic and endothelial cells. With the view of embryonic mesodermal developing events in humans, genetic modification of hES cells by lentiviral vectors provides a powerful tool for study of hematopoiesis and vasculogenesis.

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Acknowledgments

This study was partially supported by National Natural Science Foundation of China (30771093), Shanghai Pujiang Program (07pj14020) and Foundation from Science and Technology Commission of Shanghai Municipality (08DJ1400500) to Tong Chen.

Disclosures

The authors indicate no potential conflict of interest.

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Authors and Affiliations

  1. Gynecology & Obstetrics Hospital, Fudan University, Shanghai, 200011, China

    Hua Jiang, Xiaolong Lin & Youji Feng

  2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, 200040, China

    Yi Xie & Tong Chen

  3. Shanghai Ji Ai Genetics & IVF Institute, Shanghai, 200011, China

    Jinlan Han & Yueping Zhang

  4. Maine Medical Center Research Institute, Scarborough, ME, 04074, USA

    Zack Z. Wang

Authors
  1. Hua Jiang
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  2. Xiaolong Lin
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  3. Youji Feng
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  4. Yi Xie
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  5. Jinlan Han
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  6. Yueping Zhang
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  7. Zack Z. Wang
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  8. Tong Chen
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Corresponding author

Correspondence to Tong Chen.

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Hua Jiang and Xiaolong Lin contributed equally to this work.

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Jiang, H., Lin, X., Feng, Y. et al. Hemato-endothelial differentiation from lentiviral-transduced human embryonic stem cells retains durable reporter gene expression under the control of ubiquitin promoter. Cytotechnology 62, 31–42 (2010). https://doi.org/10.1007/s10616-010-9258-0

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  • DOI: https://doi.org/10.1007/s10616-010-9258-0

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