Open Access Open Badges Research article

The chick chorioallantoic membrane as an in vivo xenograft model for Burkitt lymphoma

Marcel Klingenberg1, Jürgen Becker1, Sonja Eberth2, Dieter Kube2 and Jörg Wilting1*

Author Affiliations

1 Department of Anatomy and Cell Biology, University Medical Center Goettingen, Kreuzbergring 36, Goettingen 37075, Germany

2 Department of Hematology and Oncology, University Medical Center Goettingen, Robert-Koch-Strasse 40, Goettingen 37075, Germany

For all author emails, please log on.

BMC Cancer 2014, 14:339  doi:10.1186/1471-2407-14-339

Published: 18 May 2014



Burkitt lymphoma (BL) is an aggressive malignancy that arises from B-cells and belongs to the group of Non-Hodgkin lymphomas (NHL). Due to the lack of appropriate in vivo models NHL research is mainly performed in vitro. Here, we studied the use of the chick chorioallantoic membrane (CAM) for the generation of human BL xenograft tumors, which we compared with known characteristics of the human disease.


In order to generate experimental BL tumors, we inoculated human BL2B95 and BL2-GFP cells on the CAM. BL2B95 xenograft-tumors were grown for seven days and subsequently analyzed with transmission electron and immunofluorescence microscopy, as well as histological staining approaches. BL2-GFP cells were studied at regular intervals up to seven days, and their metastatic behavior was visualized with intravital immunofluorescence techniques.


Xenografted BL2B95 cells formed solid tumors in the CAM model with a Ki67-index greater than 90%, preservation of typical tumor markers (CD10, CD19, CD20), a ‘starry sky’ morphology, production of agyrophilic fibers in the stroma, formation of blood and lymphatic vessels and lymphogenic dissemination of BL2B95 to distant sites. We identified macrophages, lymphocytes and heterophilic granulocytes (chick homolog of neutrophils) as the most abundant immune cells in the experimental tumors. BL2-GFP cells could be traced in real-time during their distribution in the CAM, and the first signs for their dissemination were visible after 2-3 days.


We show that xenografted BL2B95 cells generate tumors in the CAM with a high degree of cellular, molecular and proliferative concord with the human disease, supporting the application of the CAM model for NHL research with a focus on tumor-stroma interactions. Additionally we report that BL2-GFP cells, grafted on the CAM of ex ovo cultured chick embryos, provide a powerful tool to study lymphogenic dissemination in real-time.

Non-Hodgkin lymphoma; Angiogenesis; Lymphogenic metastasis; BL2; BL2B95; Tumor-stroma interaction; Microenvironment; Macrophages; Granulocytes