Project description:Gene expression profile of two reporgrammed cell lines iPSC CRL1831 (induced pluripotent stem cells) and CSC DLD1 (cancer stem-like cells) derived from normal colon CRL1831 and colorectal cancer DLD-1 cells, after transfer to 3D cell culture conditions and cell lines treated with single or fractionated ionizing radiation doses under 3D cell culture conditions. There are no data that cancer metastases arise due to specific mutations of cancer cells. Therefore ongoing investigation of reprogrammed cancer cells grown in three-dimensional (3D) cell culture models might provide researchers with essential data studying tumor oncogenesis and metastases formation. 3D culture models were shown to mimic in vivo cell microenvironment more accurately than the standard two-dimensional cell monolayer (2D) cultures. Also, growing evidence suggests that 2D and 3D cultured cells gene expression pattern variance following irradiation is highly dependent on cancer cell state and their interaction with microenvironment.
Project description:To investigate the differential transcriptomics upon DMT1 in 2D vs 3D cell culture in breast cancer cells Gene expression analysis from MDA-MB-231 RNA-seq data of WT and DMT1 KO cells both in 2D and 3D cell culture conditions
Project description:Tumor cell response to irradiation also depends on their microenvironment. Therefore ongoing investigation of three-dimensional (3D) cell culture models provide researchers with essential data studying and remodeling radiotherapeutic implications in cancer treatment. 3D culture models were shown to mimic in vivo cell microenvironment more accurately than the standard two-dimensional cell monolayer (2D) cultures. Growing evidence suggests that 2D and 3D cultured cell gene expression pattern discrepancies following irradiation is highly dependent on cell-ECM interactions. It has been shown that laminin-rich-extracellular matrix (lr-ECM) used in 3D cultures not only alters cancer cell phenotype and response to external stimuli but also affects their differentiation, migration and survivability. Thus, a change in these fundamental cell properties demands us to reconsider data previously collected using 2D in vitro models. RNA was harvested from two colorectal cancer cell lines cultivated under 3D cell culture conditions, 4h after treatment of single (2 Gy or 10 Gy) or fractionated (5x2 Gy) ionizing radiation dose.
Project description:Background: The main focus of the work was the evaluation of gene expression differences between our established NSCLC 3D cell culture model and the 2D cell culture in regard to the use of our model for drug screening applications. Methods: The non-small cell lung cancer (NSCLC) cell lines Colo699 and A549 were cultivated as monolayer (2D) on cell culture plates for five days or as microtissues (3D) in a hanging-drop system for five and ten days, respectively. Cells and microtissues were harvested and Affymetrix chip analyses were performed with the prior isolated RNA. This was repeated in three independent experiments. Subsequent biostatistical data analyses tested for reproducibility, comparability and significant differences in gene expression profiles between cell lines, experiments and culture methods. Results: The analyses revealed a high interassay correlation within the distinct culture systems, thus proving a high validity of our data. The comparison of 3D versus 2D cell cultures revealed significant differences in RNA expression (979 genes for A549; 1106 genes for Colo699), but the overlap of changes in RNA profiles between the cell lines at the individual gene level was small (149 genes), potentially reflecting overall heterogeneity and their origin, i.e. primary vs pleural effusion. Nevertheless, these RNA expression changes affected most relevant cancer-associated pathways as DNA methylation, cell cycle, rRNA expression and meiosis pathways. Furthermore, the expression differences between 2D and 3D were more evident after longer cultivation time, which supports the hypothesis of cultivation related mechanisms and the usage of long-time cultivation systems. Conclusion: In summary, our data support the need of innovative 3D drug testing systems to close the gap between in-vitro drug screening and in-vivo data. Thus, our 3D NSCLC model might provide a model to address the challenge of microenviroment associated resistance mechanisms, as well as cell-cell interaction related effects.
Project description:Patient-derived cancer cells (PDCs) were established by three-dimensional (3D) spheroid culture from testicular germ cell tumor (GCT) specimens. Microarray expression analysis revealed that cancer stem-like cell-related genes were upregulated in 3D culture condition compared with two-dimensional (2D) culture condition.
Project description:3D cultivation of cells lead to changes in morphology of the cells. This is likely to explain the higher radioresistance of cells growing in 3D compared to cells growing in 2D cell culture. Whole genome gene expression is performed to determine genes involved in changes of cell moroholgy and radioresistance. Keywords: comparison of 2D vs. 3D cell culture RNA of cells was isolated four days after growing in the two different cell culture systems
Project description:To better understand the tumour-modelling efficacy of cell lines, we performed RNA-seq analyses on 2D and 3D cultures of tumourigenic MCF7 and non-tumourigenic MCF10A and augmented our data with published datasets. To our knowledge, this was the first RNA-seq dataset comprising 2D and 3D cultures of MCF7 and MCF10A within the same experiment. We then compared tumourigenesis within cell lines (MCF7-vs-MCF10A) and tumourigenesis within clinical tissues (Luminal A-vs-Normal) from the TCGA-BRCA dataset and considered whether the same cancer-related processes could be observed.