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Background: cCSCs are a small subset of circulating tumor cells with cancer stem cell features: resistance to cancer treatments and the capacity for generating metastases. PDX are an appreciated tool in oncology, providing biologically meaningful models of many cancer types, and potential platforms for the development of precision oncology approaches. Commonly, mouse models are used for the in vivo assessment of potential new therapeutic targets in cancers. However, animal models are costly and time consuming. An attractive alternative to such animal experiments is the chicken chorioallantoic membrane assay. Methods: In this study, primary cultures from cCSCs were established using the sphere-forming assay. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form secondary microtumors. Results: We have developed an innovative in vitro platform for cultivation of cCSCs from peripheral blood of cancer patients. The number of tumorspheres increased significantly with tumor progression and aggressiveness of primary tumor. The number of tumorspheres was positively correlated with Ki-67, Her2 status, and grade score in primary breast tumors. The grafting of tumorspheres onto the CAM was successful and positively correlated with aggressiveness and proliferation capacity of the primary tumor. These tumors pathologically closely resembled the primary tumor. Conclusions: The number of tumorspheres cultured from peripheral blood and the success rate of establishing PDX directly reflect the aggressiveness and proliferation capacity of the primary tumor. A CAM-based PDX model using cCSC provides a fast, low-cost, easy to handle, and powerful preclinical platform for drug screening, therapy optimization, and biomarker discovery.

cancers

Chick Chorioallantoic Membrane (CAM) Assays as a Model of Patient-Derived Xenografts from Circulating Cancer Stem Cells (cCSCs) in Breast Cancer Patients