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Research Article

A rapid and simple assay for human blood malignancy engraftment, homing and chemotherapy treatment using fluorescent imaging of avian embryos

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Pages 472-478 | Received 06 May 2011, Accepted 04 Sep 2011, Published online: 15 Nov 2011
 

Abstract

Detection of grafted human cells in mice using fluorescence is a rapid and simple technique whose use is continually expanding. Robust engraftment of human hematological malignancy (HHM) lines and patient cells into the naturally immunodeficient turkey embryo has recently been demonstrated by polymerase chain reaction (PCR), fluorescence activated cell sorting (FACS) and histology. We demonstrate here that fluorescence imaging is a rapid and simple technique for detecting engraftment and homing of cells derived from HHM in turkey embryos. Raji lymphoma cells expressing a far-red fluorescent protein were injected intravascularly into turkey embryos and fluorescence was detected 8 days later in their limbs and skulls. Much stronger signals were obtained after removal of the bones from the limbs. Unlabeled Raji cells did not give a fluorescent signal. Treatment with doxorubicin dramatically reduced the fluorescent signal. Intravenously injected HL-60 leukemia cells labeled with infrared-fluorescing dye were detected in the bone marrow after 16 h. Homing was active, although some non-specific fluorescence was present. Use of fluorescence imaging of HHM in turkey embryos is therefore feasible and reduces the time, effort and expense for detecting engraftment. This technique has potential to become a high-throughput xenograft system for hematological chemotherapy development and testing, and for study of hematological cell homing.

Acknowledgements

Our deep gratitude goes to Dr. Alexander Perelman for his dedication in setting up and teaching us to use the IVIS Maestro and spectral unmixing systems. Thanks are also due to Dr. Ronen Yehuda for his operation assistance. Chaya Morgenstern provided logistic and administrative assistance. Dr. Kfir Lapid provided helpful tips and Prof. Tsvee Lapidot made a generous gift of DiR for preliminary experiments. This research was supported by a grant by the Horowitz Foundation for applied biomedical research (R.S.G.). The IVIS Maestro facility is part of the Bar-Ilan Institute of Nanotechnology and Advanced Materials.

Potential conflict of interest: Disclosure forms provided by the authors are available with the full text of this article at www.informahealthcare.com/lal.

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