At the time of diagnosis more than 65% of patients with colorectal cancer have advanced disease and require chemotherapeutic treatment. However, colorectal tumors demonstrate pronounced heterogeneity, leading to significant individual variation in sensitivity to standard chemotherapeutic drugs. Tools for assessing chemosensitivity and tailoring therapeutic regimens to the individual patient are therefore urgently needed. A promising approach is to isolate viable cancer cells from patient tissue and to test the chemosensitivity of the individual tumor in vitro.

The aim of the current study was to establish primary colorectal cancer cell cultures from patient tissue and investigate if the cultured cells retain important characteristics of the original tumor. Small cell clusters were isolated from colorectal tumor tissue and allowed to form cell spheroids in suspension culture. The cell type composition of patient spheroids was characterized by immunostaining with cell type specific antibodies. Spheroid size was monitored over time and growth curves established for each patient. The distribution of proliferating and apoptotic cells in the spheroids was assessed by staining for specific markers. The 3D culture system was verified as a tumor model by comparing the results obtained for the spheroids to paraffin-embedded sections of the original tumors.

Successful spheroid culture was obtained from more than 80% of tissue samples. Immunostaining confirmed that the primary cultures were composed of colorectal cancer cells, without significant contaminations by other cell types. Growth could be observed as an increase in spheroid size. The dominating cell populations of the original tumor could generally be identified in the derived spheroids as well. In summary, spheroid cultures of colorectal cancer cells can be established from tumor tissue of individual patients with a high success rate and the formed spheroids retain important cellular characteristics of the original tumor. Culture of colorectal cancer cells as spheroids represents a promising method for establishing primary cultures for chemotherapeutic screening of the individual patient.

Citation Format: Maria Jeppesen, Grith Hagel, Anders Glenthoj, Ole Thastrup, Lars Nannestad Jorgensen, Jacob Thastrup. Spheroid culture of colorectal cancer cells from patient tumor tissue preserves important cellular characteristics of the parental tumor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2718. doi:10.1158/1538-7445.AM2013-2718