Iris M. Krainer1, Ruth Joas1, Przemyslaw A. Filipek1, Edith Lorenz2,3, Arno Amann2,3, Lukas A. Huber1,4, Ronald Gstir1
1Austrian Drug Screening Institute, 6020 Innsbruck, Austria; 2 Medical University of Innsbruck, Department of Internal Medicine V, 6020 Innsbruck, Austria; 3 Tyrolean Cancer Research Institute, 6020 Innsbruck, Austria; 4 Medical University of Innsbruck, Biocenter, Division of Cell Biology, 6020 Innsbruck, Austria.
Publication Date 06/2019; tpub_213
Tumor derived organoids are shown from two different patients with colorectal cancer. The cultures are grown out in five different media that, to our knowledge, support the growth of most colorectal tumors. The RealTime-Glo™ MT Cell Viability Assay was started right after seeding the tumor derived cells and was analyzed over a time period of 72 hours. The growth curves established by the signal from the RealTime-Glo™ MT Cell Viability Assay recapitulated very quickly (within hours), revealing which media strategies supported the outgrowth of clones from the tumor tissue. Medium M2, M3, M4 and M5 supported tumor organoid growth from the tumor of patient 1 (P1). Medium M1 did not show satisfying growth support of tumor organoids (Fig. 1).
However, the tumor of patient 2 (P2) showed a completely different outgrowth behavior. Tumor organoids from patient P2 were ideally supported by medium M2 and M4. Medium M1 showed also acceptable growth support (Fig. 2). The cultures in medium M3 and M5 showed very weak growth and were terminated after 72 hours of culture.
Figure 1. Evaluation of organoid growth for patient P1-derived colorectal cancer organoids by RealTime-Glo™ MT Cell Viability Assay. 33,000 cancer cells/well from patient P1 were plated in a 96-well plate in 5 different media (M1, M2, M3, M4 and M5). Cell viability was monitored by RealTime-Glo™ MT Cell Viability Assay at 1, 24, 48 and 72 hours after culture start. RLU were normalized to the 1 hour value of M1. Media M2, M3, M4, and M5 supported sufficient growth whereas M1 did not support satisfying growth.