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doi:10.1038/sj/leu/2401432

Cited by 3 publications

References 6 publications

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Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

et al. 2002

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Primitive human hematopoietic cells in granulocyte-colony stimulating factor (G-CSF)-mobilized peripheral blood (MPB) are more difficult to transduce compared to cells from umbilical cord blood. Based on the hypothesis that MPB cells may require different stimulation for efficient retroviral infection, we compared several culture conditions known to induce cycling of primitive hematopoietic cells. MPB-derived CD34(+) cells were stimulated in the presence or absence of the murine fetal liver cell line AFT024 in trans-wells with G-CSF, stem cell factor (SCF), and thrombopoietin (TPO) (G/S/T; 100 ng/ml) or Flt3-L, SCF, interleukin (IL)-7, and TPO (F/S/7/T; 10-20 ng/ml), and transduced using a GaLV-pseudotyped retroviral vector expressing the enhanced green fluorescence protein (eGFP). Compared to cultures without stroma, the presence of AFT024 increased the number of transduced colony-forming cells (CFC) by 3.5-fold (with G/S/T), long-term culture-initiating cells (LTC-IC) by 4.6-fold (with F/S/7/T), and nonobese diabetic/severe immunodeficiency disease (NOD/SCID)-repopulating cells (SRC) by 6.8-fold (with F/S/7/T). Similar numbers of long-term culture-initiating cells (LTC-IC) and SRC could be transduced using AFT024-conditioned medium (AFT-CM) or a defined medium that had been supplemented with factors identified in AFT-CM. Finally, using our best condition based on transduction with the gibbon ape leukemia virus (GaLV)-pseudotyped vector, we demonstrate a 33-fold higher level of gene transfer (p < 0.001) in SRC using an RD114-pseudotyped vector. In summary, using an optimized protocol with low doses of cytokines, and transduction with an RD114 compared to a GaLV-pseudotyped retroviral vector, the overall number of transduced cells in NOD/SCID mice could be improved 144-fold, with a gene-transfer efficiency in SRC of 16.3% (13.3-19.9; n = 6).

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Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

et al. 2002

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Bone Morphogenetic Protein 4 Contributes to the Maintenance of Primitive Cord Blood Hematopoietic Progenitors in an Ex Vivo Stroma-Noncontact Co-Culture System

Hutton

Rozenkov

Khor

et al. 2006

Stem Cells and Development

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Establishment of conditions supporting hematopoietic stem cell (HSC) maintenance and expansion ex vivo is critical for wider clinical application of cord blood (CB) transplantation. AFT024 is a murine fetal liver cell line that expands primitive hematopoietic cells via a process that is not understood. Here we show that bone morphogenic protein 4 (BMP4) is produced by AFT024 and contributes significantly to the maintenance of co-cultured CB-derived primitive cells. Significant amounts of BMP4 mRNA are produced by the supportive AFT024 stromal cell line, and secreted BMP4 protein accumulates in AFT024 conditioned medium. Blockade of BMP4 activity in this coculture model using neutralizing BMP4 monoclonal antibody reduced expansion of primitive CB cells on the basis of phenotypic (CD34(+)CD38(-)) and functional criteria [long-term culture initiating cells (LTC-IC)] and significantly reduced the capacity of the cultured CB stem cells to support repopulation in the nonobese diabetic-severe combined immunodeficiency (NOD-SCID) xenograft model. Therefore, BMP4 is a key growth factor for maintenance of HSC and contributes to the unique properties of the AFT024 stromal noncontact culture.

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Efficient generation of CAR NK cells from human umbilical cord blood CD34⁺stem and progenitors for democratizing affordable immunotherapy

Li,

Wang,

Zheng

et al. 2024

Preprint

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Chimeric antigen receptor (CAR) natural killer cells (CAR NK) cells, leveraging safety and not requiring HLA match in adoptive infusion, have emerged as promising alternative cells to CAR-T cells for immunotherapies. High and multiple doses of CAR NK cell infusions are essential to maintain therapeutic efficacy in clinical trials. This requires efficient methods for generating large-scale CAR NK cells and significantly reducing CAR engineering costs. In this study, we develop a three-step strategy to generate highly high yields of induced NK (iNK) and CAR iNK cells from human umbilical cord blood CD34+hematopoietic stem and progenitor cells (CD34+HSPCs). Starting from a single umbilical cord blood CD34+HSPC, our reliable method efficiently produces 14-83 million mature iNK cells or 7-32 million CAR iNK cells with high expression levels of CD16 and zero T cell contaminations. Introducing CAR expression elements at the HSPC level reduces the quantities of CAR pseudoviruses to 1 / 140.000 - 1 / 600,000 compared to engineering CARs in mature NK cells. The iNK and CAR iNK cells, including fresh cells and thawed cells from cryopreserved conditions, demonstrate remarkable tumoricidal activities against various human cancer cells and significantly prolong the survival of human tumor-bearing animals. The high yields of CAR NK cells and negligible costs of CAR engineering of our method support the broad applications of CAR NK cells for treating cancer patients.

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Cited by 3 publications

References 6 publications

Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

Bone Morphogenetic Protein 4 Contributes to the Maintenance of Primitive Cord Blood Hematopoietic Progenitors in an Ex Vivo Stroma-Noncontact Co-Culture System

Efficient generation of CAR NK cells from human umbilical cord blood CD34⁺stem and progenitors for democratizing affordable immunotherapy

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Cited by 3 publications

References 6 publications

Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

Optimization of Gene Transfer into Primitive Human Hematopoietic Cells of Granulocyte-Colony Stimulating Factor–Mobilized Peripheral Blood Using Low-Dose Cytokines and Comparison of a Gibbon Ape Leukemia Virus Versus an RD114-Pseudotyped Retroviral Vector

Bone Morphogenetic Protein 4 Contributes to the Maintenance of Primitive Cord Blood Hematopoietic Progenitors in an Ex Vivo Stroma-Noncontact Co-Culture System

Efficient generation of CAR NK cells from human umbilical cord blood CD34+stem and progenitors for democratizing affordable immunotherapy

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Efficient generation of CAR NK cells from human umbilical cord blood CD34⁺stem and progenitors for democratizing affordable immunotherapy