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Invasion, proliferation and influence of cancer cell line MDA-MB-231 on organotypic tissue using Precision Cut Lung Slices (PCLS)

: Konzok, Sebastian
: Braun, Armin; Tamura-Niemann, Teruko

Hannover, 2014, IV, 82 pp.
Hannover, Medizinische Hochschule, Master Thesis, 2014
Master Thesis
Fraunhofer ITEM ()
precision cut lung slices (PCLS); cancer cell proliferation ex vivo; breast - cancer; Cell lines

Cancer is a major public health problem with breast cancer being among the leading causes of cancer mortality worldwide. The actual cause of death is the formation of metastasis which often occurs in the lung. Among the cell types present in the tumor microenvironment, macrophages have proven to be the dominant leukocyte population. An increasing amount of publications points to evidence that a macrophage sub-population actively supports and promotes the initiation, growth and development of tumor tissue. Indeed, Bingle et al. [2002] have shown in their meta-analysis that over 80% of studies show a correlation between macrophage density and poor patient prognosis. As a result, the local immune response to tumor initiation is a valuable research topic and the development of models for testing of cancer therapeutics is highly sought after. To this end, we utilize an innovative organotypic tumor invasion model, using living human precision-cut-lung-slices (PCLS) and cancer cells in an effort to focus on the local immunological reactions of the tissue during the first stages of metastasis formation. An AdGFP-transduced human breast cancer cell line MDA-MB-231 was added to human PCLS over a period of one week. Viability assays such as LIVE/DEAD staining and LDH measurements were performed to assure intact human tissue. Tissue immune staining against CD68 was used to visualize locations of macrophage-MDA-MB-231 interaction whereas a staining against Ki67 as a proliferation marker was performed to visualize proliferating cells. Immune response and neoangiogenesis were determined by cytokines IL-10 and IL-1beta, angiogenesis marker VEGF and tumor marker GM-CSF, respectively. Human Precision Cut Lung Slices (PCLS) remain viable during infection and invasion with cancer cells. Cancer cells expand and proliferate in human lung tissue, which could be shown through quantification of the Ki67- positive MDA- MB- 231 cells as well as videos of the dynamic changes within the tissue. CD78- positive cells were found to be associated with MDA-MB-231 cells as colo-calization of CD68- positive macrophages and MDA-MB-231 was found during the entire invasion period. The VEGF/GM-CSF release correlated with the MDA-MB-231 growth curves whereas IL- 1beta showed a strong pro-inflammatory signal 5 hours after seeding and a continuous decline thereafter. First experiments to adapt the model to different species such as mice were performed that show that the cancer cells also attach themselves to the tissue matrix of other species. Here we mimic cancer cell proliferation in the microenvironment of human lung tissue without and thus provide insights into functional local immune responses with a human physiology background.