I. Introduction
Recently, the US FDA has approved Novartis's CAR-T therapy, Kymriah, and the prospect of cancer immunotherapy is broad. However, the current direction of most immunotherapy is still focused on T cell regulation: either through T cell gene editing, such as CAR-T treatment; or through immune checkpoint inhibition, restore T cell killing ability to cancer cells, Kymriah's listing And recent clinical success stories have greatly supported research in this area. However, we can also study immune cells and their functions in many other ways. For example , the function and application of macrophages in tumor cell clearance has received much attention.
Second, the lecture introduction
This lecture describes the use of IncuCyte "Viable Cell Dynamic Imaging and Analysis System" , combined with IncuCyte "pHrodo Red Cell Labeling Kit" to observe dynamic live cell phagocytosis experiments, including the principle of cell labeling, the effect of drugs on "phagocytosis", experimental verification And supplemental phenotypic experiments. Anti-CD47 antibody was demonstrated as a potential novel immunotherapeutic drug that mediates the phagocytosis of cancer cells by macrophages.
Lecture title
Third, the experimental principle
IncuCyte "pHrodo Red Cell Labeling Kit" can label target cells with pH-sensitive probes . When these cells are in culture, the probes do not produce fluorescence. However, when they are phagocytosed by phagocytic cells, they enter a lower pH phagolysosome, which can significantly increase the fluorescence of the probe. The video on the right shows that larger elongated macrophages are phagocytizing smaller target cells. After the target cells are phagocytosized, the red fluorescence in the macrophages is enhanced and the IncuCyte live cell dynamic imaging is built into the incubator. The Analytical System detects the number of cells that emit red fluorescence and performs dynamic imaging and data analysis of the phagocytic process.
The principle of "phagocytosis experiment"
Fourth, the experimental results
Anti-CD47 antibody-mediated "cell phagocytosis" image
Curve of fluorescence object over time during anti-CD47 antibody-mediated "phagocytosis"
Quantification of "cell phagocytosis": inhibition of "phagocytosis" by four drugs
V. Automated live cell dynamic analysis
At present, most of the cell detection methods still belong to the traditional end point method - only the final result is given. This method can not only dynamically monitor and analyze the whole process of cell experiments, but also obtain biased results. In order to be able to observe live cells throughout the process, Essen has developed the IncuCyte “Viable Cell Dynamic Imaging and Analysis System†built into the incubator, inside the incubator, and simultaneously on 6 cell culture dishes/bottles/microplates (6- 384-well) for long-term dynamic imaging, recording real-time changes in cell morphology details, and transforming into a variety of cell-related time-quantitative curves through powerful image analysis software.
IncuCyte S3
IncuCyte is a cell-level scientific and drug screening solution that has been used for immunosuppression, nerve growth and regression, cancer cell metastasis, cell migration, stem cell differentiation, apoptosis, cell necrosis, cell proliferation, wound repair, etc. Scale screening and research on long-term life science issues.
Sixth, online lecture video
Video length is 27min, please watch under WiFi conditions:
Https://v.qq.com/x/page/k0538aoejcj.html
Seventh, about "classic creatures"
"Diaoao Bio" (Shanghai, Hong Kong, Taiwan) was established in 1994. For more than 20 years, it has been adhering to the world-class instrument manufacturers. The experts with rich R&D experience are technical support, and the professional maintenance team is backing the service. Focus on providing the most advanced hardware and software equipment and technical services for life sciences, pharmaceutical research and development and disease treatment.
The instruments of the exclusive distributor of "Diaoao Bio" are:
IncuCyte, Beacon, Gyrolab, iQue Screener, Avatar, PlexBio, Certus, HighRes, etc.
Eight, contact information
"Shanghai Dianao Biotechnology Co., Ltd." :
Website:
Tel: 021-58605185 (Transfer Department)
Email:
Scan the QR code to add the WeChat public account "tekontech" :
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