Abstract
Ewing tumors are a clinically heterogeneous group of childhood sarcomas that represent a paradigm for understanding solid tumor biology, as they are the first group of sarcomas for which a chromosome translocation has been characterized at the molecular level. However, the biologic organization of the tumor, especially the processes that govern proliferation, differentiation, and metastasis of primitive tumor stem cells is poorly understood. Therefore, to develop a biologically relevant in vivo model, five different Ewing tumor cell lines and primary tumor cells from three patients were transplanted into immune-deficient mice via intravenous injection. NOD/ scid mice that carry a complex immune deficiency and thus nearly completely lack the ability to reject human cells were used as recipients. Overall, 26 of 52 mice (50%) transplanted with VH-64, WE-68, CADO-ES1, TC-71, and RM-82 cells and 4 of 10 mice (40%) transplanted with primary tumor cells engrafted. Moreover, primary cells that did not grow in vitro proliferated in mice. The pattern of metastasis was similar to that in patients with frequent metastases in lungs (62%), bone marrow (30%), and bone (23%). Using limiting dilution experiments, the frequency of the engraftment unit was estimated at 1 Ewing tumor-initiating cell in 3 × 105 VH-64 cells. These data demonstrate that we have been able to establish an in vivo model that recapitulates many aspects of growth and progression of human Ewing tumors. For the first time, this model provides the opportunity to identify and characterize primitive in vivo clonogenic solid tumor stem cells. This model will, therefore, be instrumental in studying many aspects of tumor cell biology, including organ-selective metastasis and tumor angiogenesis.
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Abbreviations
- APAAP method:
-
alkaline phosphatase anti-alkaline phosphatase method
- CyChrome:
-
phycoerythrin-cyanine
- DTAF:
-
dichlorotrianzinylamino-fluorescein
- ET-IC:
-
Ewing tumor-initiating cell
- H&E:
-
hematoxylin and eosin
- HLA:
-
human leukocyte antigen
- NOD/ scid mice:
-
nonobese diabetes/severe combined immune-deficiency mice
- PAS:
-
periodic acid-Schiff reaction
- PE:
-
phycoerythrin
- PNET:
-
peripheral neuroectodermal tumor
- VCAM:
-
vascular adhesion molecule
- VLA:
-
very late activation antigen
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Acknowledgements
The authors thank Prof. J.E. Dick, Toronto, Ontario, Canada, Prof. A.L. Harris, O-ford, U.K., and Prof. J. Kienast and Dr. M. Paulussen, Münster, Germany, for stimulating discussions and for critically reviewing the manuscript, and Prof. K.-D. Richter, Central Animal Care Facility, Münster, Germany, for his continuing support. We are also grateful to our technicians Andrea Hötte, Frauke Schmidt, and Tanja Möllers for technical help.
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Supported by grants #10-0995-Vo 1 and #10-1252-Vo 2 from the Deutsche Krebshilfe.
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Vormoor, J., Baersch, G., Decker, S. et al. Establishment of an In Vivo Model for Pediatric Ewing Tumors by Transplantation into NOD/ scid Mice. Pediatr Res 49, 332–341 (2001). https://doi.org/10.1203/00006450-200103000-00006
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DOI: https://doi.org/10.1203/00006450-200103000-00006
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