Stejskalová, Anna; Fincke, Victoria; Nowak, Melissa K.; Nowak, Melissa Kim; Nowak, Melissa; Schmidt, Yvonne; Borrmann, Katrin; Wahlde, Marie‑Kristin von; Schäfer, Sebastian Daniel; Kiesel, Ludwig; Greve, Burkhard; Götte, Martin: Collagen I triggers directional migration, invasion and matrix remodeling of stroma cells in a 3D spheroid model of endometriosis
Inhalt
- An endometrial stromal cell line (St-T1b), primary endometriotic stromal cells and the endometriotic epithelial cell line (12Z) self-organize into spheroids in hanging drop culture.
- Matrigel and collagen I trigger distinct phenotypes in single cells and spheroids where stromal condensates create defects on collagen I.
- Directional invasion followed by the formation of a circular defect occurs in St-T1b and ESCs spheroids but not in St-T1b:12Z co-culture.
- Spheroid 3D culture as an effective tool to screen small molecule drug and microRNA-based therapeutics.
- The broad-spectrum MMP inhibitor NNGH limits the invasive behaviour of stromal spheroids on collagen I.
- ROCK inhibition significantly enhances spreading and invasion of all studied endometrial cell types on Matrigel.
- The spheroid model reveals context-dependent roles of the mechanoregulatory microRNAs miR-200b and miR-145 on the invasive behaviour of the endometriotic epithelial cell line 12Z on Matrigel.
- Discussion
- Methods
- Cell culture.
- Spheroid formation.
- Preparation of collagen I and Matrigel.
- Spheroid response to collagen IMatrigel.
- Metabolic activity measurement.
- Inhibitors.
- microRNA transfection.
- Live cell staining and immunostaining.
- Imaging.
- Image analysis.
- RNA extraction and cDNA synthesis.
- PCR.
- Statistical analysis.
- References
- Acknowledgements