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Stable and reproducible engraftment of primary adult and pediatric acute myeloid leukemia in NSG mice

Leukemia volume 25, pages 1635–1639 (2011)Cite this article

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Models of primary human acute myeloid leukemia (AML) are important tools to study various aspects of the pathophysiology of AML, like stem cell biology, multimodal drug therapies and development of resistance in vivo. In contrast to acute lymphoblastic leukemia, AML xenotransplant models are hampered by persistent problems of inadequate engraftment and a short-life span of the animals. Increasingly, immunodeficient strains evolved from the NOD/SCID strain to evade residual innate immunity (summarized in Shultz et al.1). The NOD.Cg-PrkdcScidIL2 rgtm1Wjll/SzJ (NSG) mouse model is the product of a targeted deletion of the common gamma-chain and represents a substantial improvement leading to a complete and stable deficiency of mature lymphocytes and NK cells, a prolonged survival beyond 16 months of age and an acceptable radiation sensitivity highly permissive for engraftment of primary human cells.2 Besides the animal strains, a variety of techniques are described to transplant human AML samples. Most of the previous studies published engraftment rates after intravenous infusion between 37.8 and 70% with a median delay from 8 weeks to 12 weeks.3, 4, 5 The reported results are based on intramedullary injections to transplant human AML cells. We demonstrate a mean engraftment of 81.6% with a median delay of 10 weeks.

Successful serial transplants up to the fifth generation are currently performed with 2 × 106 unsorted cells with a mean of 1.5 × 105 hCD33+ cells in a volume of 50 μl FCS from fresh or frozen samples derived from the liver, spleen or BM. Overall, 63.9% engrafted successfully. The median delay of engraftment was reduced from 10 (3–23) weeks in the primary transplant to 9 (2–18) weeks in the secondary transplants. It seemed that engraftment delay diminishes with passage (preliminary data: mean delay for the third transplant: 4 weeks and for the fourth: 2 weeks). In secondary transplants, fresh samples engrafted in 75% with an overall median delay of 4 (2–6) weeks, whereas frozen samples had a median time to engraftment of 13 (6–19) weeks and were positive in 42.9% of the cases. Due to the single AML sample derived from PB, we were not able to compare the two different types of cells of origin. Sanchez et al.5 reported successful serial transplantations with 2.5 × 106 to 10 × 106 unsorted hCD45+/hCD33+ cells using sublethally irradiated NSG mice, which is consistent with engraftment of leukemia initiating cells (SL-IC). We can confirm these results in non-irradiated animals with substantially less hCD45+/hCD33+ cells. In addition, besides BM and spleen, hCD45+/hCD33+ cells derived from the liver of primary transplanted animals engrafted well. In our hands, serial transplants also showed a significantly reduced delay to engraftment and were not influenced by cells of origin or amount of injected cells. The repopulation capacity of these SL-IC seems to increase with the passages.

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Acknowledgements

We thank the members of the laboratory for Cytogenetic and Molecular Genetics at the Department of Internal Medicine III for their support with the primary adult samples, the AML–BFM group for providing the clinical information and patient data and all members of the animal facility of the University of Ulm. The technical assistance of Beate Junk is greatly appreciated. This work was supported by a grant from the Deutsche José Carreras Leukämie-Stiftung e.V. (DJCLS R07/23) to S Corbacioglu. All animal experiments were conducted according to 54-2532.1-27/09 approved protocol.

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Author notes

  1. M Malaisé and M Neumeier: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatrics, University of Regensburg, Regensburg, Germany

    M Malaisé, M Neumeier, C Botteron & S Corbacioglu

  2. Department of Internal Medicine III, University Hospital of Ulm, Ulm, Germany

    K Döhner

  3. Department of Pediatrics, University of Hannover, Hannover, Germany

    D Reinhardt

  4. Institute of Cell and Molecular Pathology, Hannover Medical School, Hannover, Germany

    B Schlegelberger & G Göhring

  5. Department of Pediatrics, University Children's Hospital Jena, University of Jena, Jena, Germany

    B Gruhn

  6. Department of Pediatrics, University Hospital of Ulm, Ulm, Germany

    K-M Debatin

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  1. M Malaisé
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  2. M Neumeier
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Correspondence to S Corbacioglu.

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Malaisé, M., Neumeier, M., Botteron, C. et al. Stable and reproducible engraftment of primary adult and pediatric acute myeloid leukemia in NSG mice. Leukemia 25, 1635–1639 (2011). https://doi.org/10.1038/leu.2011.121

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