Establishment and Culture of Patient-Derived Primary Medulloblastoma Cell Lines

Established cell lines have been extensively used in cancer research. They are easy to obtain and expand and are composed of a relatively uniform population of cells. When experimental conditions are kept standard, these cells allow a high reproducibility of experimental findings from independent research groups. However, because these cell lines have been propagated in culture for decades, additional genetic lesions may be acquired leading to modification of their characteristics as compared to the original tumor. Primary cultures represent a valid alternative. Here, we describe standardized protocols to establish medulloblastoma (MB) patient-derived primary cultures from fresh tumor samples. MB primary cells grow as an adherent culture on a laminin coating and can be propagated in vitro for a limited number of passages, therefore reducing the chances to accumulate molecular alterations compared to long-term cultures. Consequently, they better resemble the original tumor both in terms of biological behavior and molecular characteristics. Low-passage MB primary cells can be used as an in vitro model for biochemical studies and functional assays, representing a useful tool to dissect the contribution of molecular pathways to MB pathogenesis. They can also represent a useful screening tool for potential therapeutic agents in preclinical studies.