It is always important to choose appropriate anticancer drugs for cancer patients. At RCL, a division of Uppsala university hospital, drug resistance profiles of patients are evaluated by a cell viability assay called FMCA. However, the number of anticancer drugs that can be evaluated by the FMCA is dependent on the number of viable cancer cells from tissues that can be obtained from each individual patient. Therefore, improvement of cell viability methods is an important issue at RCL. This study was performed to improve the FMCA method by organoid culture from colorectal cancer, renal cancer and osteosarcoma to increase the number of cancer cells. As results, it was successful to expand cryopreserved patient cancer cells to organoids to acquire more cells than before expansion. Organoids showed rounded structure in microscopy images. Thereafter, FMCA was performed on organoids as well as on thawed cryopreserved cancer cells from the original sample. Those results showed that original cancer cells, cryopreserved original cancer cells and expanded organoids derived from those cryopreserved cells had similar resistance profiles. It was also discovered that the organoids secreted VEGF under the cultivation. From those results, it can be concluded that organoids are representative of the original cancer from the patients. It is however needed to improve organoid culture methods, and to further confirm organoids by protein expression analysis and DNA analysis.

Background: Cancer is a common disease, and the choice of treatment becomes more difficult over time due to chemotherapy resistant in cancer cells. To improve the in vitroassay and the individual cancer treatment, a luminescence-based endpoint assay, CellTiter Glo 2.0 was compared with the currently in use fluorescence endpoint assay, fluorometric microculture cytotoxic assay.  Aim: The aim of this study was to validate and compare the CellTiter Glo 2.0 assay with awell-established method (FMCA) and MTT [3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2Htetrazolium bromide] assay. Moreover, investigate whether the generated data can be used as a reference database for validation of patient samples in the future.   Materials and methods: The validation was performed on peripheral blood mononuclear cells from different healthy donors and two cell lines (HCT116-wt and HT-29) of colorectal cancer carcinoma were ordered frozen from American Type Culture Collection. Analysis was also done in solid samples (ovarian and kidney cancer cells). To get as correct evaluation as possible all materials were analyzed in parallel between the two methods.  Results and conclusion: A clear trend was observed when using CellTiter Glo 2.0 assay,post FMCA directly on tumor cells. This setup, makes it possible to collect reference data in the future. In addition, a high spread of the survival index data was noted between the two methods. The reason is still unknown but could be due to the low number of tested tumor cells, therefore more tumor cells need to be tested in future studies