Optimisé de gratter Assay for In Vitro test de Migration cellulaire avec une caméra optique automatisée
Please note that all translations are automatically generated.
Click here for the English version.
Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| oCelloScope | BioSense Solution ApS | Optical camera | |
| UniExplorer software | BioSense Solution ApS | (8.0.0.7144 (RL3)) | |
| HaCaT cell | Donated from Bispebjerg Hospital | ||
| DMEM (1x) + GlutaMAX | Gibco | 31966-021 | Medium for HaCaT |
| FBS | Gibco | 10270 | Fetal bovine serum |
| PBS | Gibco | D837 | Without Mg+ and Ca2+ |
| Pencillin-Streptomycin | Sigma | P0781 | Antibiotics |
| Trypsin (10x) | Sigma | T3924 | |
| Mitomycin C | Tocris | 3258 | Inhibits proliferation |
| Microtiter plate | Greiner Bio-one | 677 180 | |
| Incubator | Panasonic | 37°C, 5% CO2 | |
| Hemocytometer | Assistent | Türk (0.1 mm) | |
| Pipet boy | Accu-Jet pro |
References
- Frykberg, R. G., Banks, J. Challenges in the Treatment of Chronic Wounds. Advances in wound care. 4 (9), 560-582 (2015).
- da Silva, L., Carvalho, E., Cruz, M. T. Role of neuropeptides in skin inflammation and its involvement in diabetic wound healing. Expert opinion on biological therapy. 10 (10), 1427-1439 (2010).
- Vyas, K. S., Wong, L. K. Detection of Biofilm in Wounds as an Early Indicator for Risk for Tissue Infection and Wound Chronicity. Annals of Plastic Surgery. 76 (1), 127-131 (2016).
- Paul, C. D., Mistriotis, P., Konstantopoulos, K. Cancer cell motility: lessons from migration in confined spaces. Nature Reviews Cancer. 17 (2), 131-140 (2016).
- Folkman, J. Angiogenesis. Annual review of medicine. 57, 1-18 (2006).
- Monsuur, H. N., et al. Methods to study differences in cell mobility during skin wound healing in vitro. Journal of Biomechanics. 49 (8), 1381-1387 (2016).
- Tang, L., et al. Human lactoferrin stimulates skin keratinocyte function and wound re-epithelialization. The British journal of dermatology. 163 (1), 38-47 (2010).
- Ashby, W. J., Zijlstra, A. Established and novel methods of interrogating two-dimensional cell migration. Integrative Biology. 4 (11), 1338-1350 (2012).
- Klettner, A., Tahmaz, N., Dithmer, M., Richert, E., Roider, J. Effects of aflibercept on primary RPE cells: toxicity, wound healing, uptake and phagocytosis. British Journal of Ophthalmology. 98 (10), 1448-1452 (2014).
- Doyle, W., Shide, E., Thapa, S., Chandrasekaran, V. The effects of energy beverages on cultured cells. Food and Chemical Toxicology. 50 (10), 3759-3768 (2012).
- Zordan, M. D., Mill, C. P., Riese, D. J., Leary, J. F. A high throughput, interactive imaging, bright-field wound healing assay. Cytometry Part A. 79 (3), 227-232 (2011).
- Keese, C. R., Wegener, J., Walker, S. R., Giaever, I. Electrical wound-healing assay for cells in vitro. Proceedings of the National Academy of Sciences. 101 (6), 1554-1559 (2004).
- Liang, C. C., Park, A. Y., Guan, J. L. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nature Protocols. 2 (2), 329-333 (2007).
- Räsänen, K., Vaheri, A. TGF-beta1 causes epithelial-mesenchymal transition in HaCaT derivatives, but induces expression of COX-2 and migration only in benign, not in malignant keratinocytes. Journal of Dermatological Science. 58 (2), 97-104 (2010).
- Tochio, T., Tanaka, H., Nakata, S., Hosoya, H. Fructose-1,6-bisphosphate aldolase A is involved in HaCaT cell migration by inducing lamellipodia formation. Journal of Dermatological Science. 58 (2), 123-129 (2010).
- Tomasz, M. Mitomycin C: small, fast and deadly (but very selective). Chemistry and Biology. 2 (9), 575-579 (1995).
- Stamm, A., Reimers, K., Strauß, S., Vogt, P., Scheper, T., Pepelanova, I. In vitro wound healing assays – state of the art. BioNanoMaterials. 17 (1-2), 79-87 (2016).
- Justus, C. R., Leffler, N., Ruiz-Echevarria, M., Yang, L. V. In vitro Cell Migration and Invasion Assays. Journal of Visualized Experiments. (88), 1-8 (2014).
- Jonkman, J. E. N., et al. An introduction to the wound healing assay using live-cell microscopy. Cell Adhesion & Migration. 8 (5), 440-451 (2016).
- Hong, J., Kandasamy, K., Marimuthu, M., Choi, C. S., Kim, S. Electrical cell-substrate impedance sensing as a non-invasive tool for cancer cell study. The Analyst. 136 (2), 237-245 (2011).
- Fredborg, M., et al. Real-time optical antimicrobial susceptibility testing. Journal of Clinical Microbiology. 51 (7), 2047-2053 (2013).
- Jenssen, H. Anti herpes simplex virus activity of lactoferrin/lactoferricin – An example of antiviral activity of antimicrobial protein/peptide. Cellular and Molecular Life Sciences. 62 (24), 3002-3013 (2005).
- Jenssen, H., Hancock, R. E. W. Antimicrobial properties of lactoferrin. Biochimie. 91 (1), 19-29 (2009).
- Delva, E., Tucker, D. K., Kowalczyk, A. P. The desmosome. Cold Spring Harbor perspectives in biology. 1 (2), 1-17 (2009).
- Ponti, A. Two Distinct Actin Networks Drive the Protrusion of Migrating Cells. Science. 305 (5691), 1782-1786 (2004).
- Canali, C., Spillum, E., Valvik, M., Agersnap, N., Olesen, T. . Whitepaper a Digital Time-Lapse Bright Field Technology To Drive Faster, Higher Throughput Bioanalysis. , (2016).
- Peplow, P. V., Chatterjee, M. P. A review of the influence of growth factors and cytokines in in vitro human keratinocyte migration. Cytokine. 62 (1), 1-21 (2013).
- Ilina, O., Friedl, P. Mechanisms of collective cell migration at a glance. Journal of Cell Science. 122 (18), 3203-3208 (2009).
Cite This Article
Vang Mouritzen, M., Jenssen, H. Optimized Scratch Assay for In Vitro Testing of Cell Migration with an Automated Optical Camera. J. Vis. Exp. (138), e57691, doi:10.3791/57691 (2018).