Engraftment of genetically modified human amniotic fluid-derived progenitor cells to produce coagulation factor IX after in utero transplantation in mice

Authors

  • Chen-Min Yang,

    1. Laboratory of Development Biology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Department of Obstetrics and Gynecology Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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  • Xiu-Li Gong,

    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
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  • Jin Qiu,

    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
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  • Hui-Xiang Tang,

    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
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  • Zhi-Juan Gong,

    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
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  • Shu-Zhen Huang,

    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
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  • Fanyi Zeng

    Corresponding author
    1. Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
    2. Minister of Health Key Laboratory of Embryo Molecular Biology and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, China
    • Laboratory of Development Biology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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Corresponding author: e-mail: fzeng11@gmail.com

Abstract

Human amniotic fluid derived progenitor cells (hAFPCs) may be multipotent and can be considered a potential tool in the field of cell therapy for haemophilia B. Their capacity to express human coagulation factor IX (hFIX) after transduction and their fate after in utero transplantation is unknown. hAFPCs isolated from second trimester pregnancies were assessed for their phenotypic markers, multilineage capacity, and expression of hFIX after transduction. Their engraftment potential was analysed in a mouse model after in utero transplantation at embryonic day 12.5. Immunohistochemistry, fluorescence in situ, ELISA and PCR were used to assess post-transplant chimeras. hAFPCs expressed several pluripotent markers, including NANOG, SOX2, SSEA4 and TRA-1-60, and could differentiate into adipocytes and osteocytes. In vitro, after transduction with hFIX and EGFP cDNAs, constitutive hFIX protein expression and clotting activity were found. Engraftment was achieved in various foetal tissues after in utero transplantation. Safe engraftment without oncogenesis was confirmed, with low donor cell levels, but persistent engraftment, into different organs (liver, heart and lung) through to 12 weeks of age. Transgenic expression of circulating hFIX was detected in recipient mice for up to 12 weeks. hAFPCs can be engrafted long-term in immunocompetent mice after in utero transplantation. Thus, cell transplantation approaches using genetically engineered hAFPCs may prove valuable for the prenatal treatment for haemophilia B.

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