Novel alginate three-dimensional static and rotating culture systems for effective ex vivo amplification of human cord blood hematopoietic stem cells and in vivo functional analysis of amplified cells in NOD/SCID mice

Authors

  • Yan Yuan,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Wai-Yee Sin,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Bofu Xue,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Yan Ke,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Kai-Tai Tse,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Zi Chen,

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • Yi Xie,

    Corresponding author
    1. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    2. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
    • Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
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  • Yong Xie

    1. Division of Life Science, Hong Kong University of Science and Technology (HKUST), Hong Kong SAR, China
    2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China
    3. Department of Obstetrics and Gynaecology, Queen Elizabeth Hospital and BioRx Limited, HKUST, Hong Kong SAR, China
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  • This work was supported in part by Grant 81100385 and Grant 81170491 from the National Nature Science Foundation of China.

Address reprint requests to: Yong Xie, PhD, Division of Life Science, Hong Kong University of Science and Technology, Clearwater Bay, Kowloon, Hong Kong SAR, China; e-mail: boyxie@ust.hk.

Abstract

Background

Autologous and allogeneic hematopoietic stem cell (HSC) transplantations serve as effective therapy for a variety of hematologic and other diseases. Umbilical cord blood (UCB) is an important source of HSCs. However, it is difficult to obtain a sufficient number of HSCs with complete self-renewal capability derived from a single unit of UCB for use in adult transplantation. In this study, we investigated two novel three-dimensional (3D) culture systems (static and rotating) for ex vivo expansion of HSCs from UCB.

Study Design and Methods

We encapsulated the human cord blood mononuclear cells (CBMCs) in alginate 3D static and rotating culture systems, compared the cell number amplification, the proportion of CD34+ cells, and the colony-forming capacity of these systems to those of the conventional two-dimensional (2D) system. The amplified cells were transplanted into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice to confirm the hematopoiesis reconstruction capacity of the cells.

Results

The increase in the cell number and the proportion of CD34+ cells in the CBMCs was more effective in these 3D alginate culture systems than in the conventional 2D culture system under the same conditions (p < 0.05). The stem cell maintenance capability was confirmed by flow cytometry and colony-forming assay ex vivo and NOD/SCID mice xenogeneic transplantation model in vivo.

Conclusion

Our results demonstrated that these 3D alginate culture systems are an efficient way to amplify cord blood HSCs for extended periods without having them lose their self-renewal capacity in vivo. These novel 3D alginate culture systems are promising for the amplification of UCB-derived HSCs for clinical application in the future.

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