Descriptive analysis of tumor cells with stem like phenotypes in metastatic and benign adrenal tumors

Background: Neuroblastoma (NB) comprises 7% of all childhood cancers. Here we report a descriptive analysis of key cellular markers that have "stem-like" properties which theoretically represents the self-renewing population of cells responsible for generating new tumor cells. Samples are obtained from freshly isolated tissue from nonmetastatic NB, metastatic NB, benign adrenal adenoma and a ganglioneuroma. In addition, in metastatic NB, descriptive analysis of the tumor cells after 3D culture as well as reanalysis of fresh tumor obtained after surgical excision posttreatment was performed.

Methods: Cells were isolated from primary tissue and characterized via immunohistochemistry and flow cytometry for markers associated with stem-like properties. In two patients, reanalysis was performed in freshly isolated tissue after chemotherapy. In three patients, freshly isolated tumors were cultured in 3 dimensions for 7-10 days and changes in stem-like marker expression were characterized.

Results: Flow analysis of metastatic NB revealed elevated levels of markers CD133, CD24, CD44, Oct4, CXCR4 and Nestin. In addition, some markers such as CD133 and CXCR4 maintained increased expression after chemotherapy.

Conclusions: The expression profile of cells with "stem-like" properties has individual variability and differs depending on the tumor type. In metastatic NB, expression of "stem-like" markers Nestin, Oct4, and CXCR4 are maintained in a higher percentage of cells and this persists even after chemotherapy. In addition, culture of freshly isolated tissue maintained the individual expression profile of stem-like markers for at least 7 days.