From 8th to 26th of November we had a chance to participate in the first practical Science and Technology (S&T) course, organized by Università degli Studi di Milano (UMIL). The first week was combined with online theoretical lessons, in which everyone could take part. Following the introduction to immunotoxicity, the course focused on presenting and discussing the application and interpretation of in vitro immunotoxicity tests, mainly involving immunosuppression and the use of the whole blood and PBMC assay.
For the first week, the training embraced the treatment of defrost peripheral blood mononuclear cell (PBMC) and Cyclosporin A cytotoxicity evaluation with a flow cytometer analysis. We learned how to start cell culture from PBMCs, how to treat them with immunotoxic compounds in combination with immunostimulant such as PHA, and how to assess cell viability on a flow cytometer using Propidium Iodide (PI).
For the second week, the lab activity included PBMC extraction from buffy coats of healthy donors. This part of the experiment required special pipetting skills and a lot of patience. Successively isolated cells and the whole blood from the same donor were the immunotoxicity treated like in a previous week.
For the last week of the course, we performed ELISA assay, analysing hIL-8 for both experiments (PBMC and whole blood). The course finished with an analysis of the obtained results and a summary of the acquired knowledge.
It was a pleasure to be able to attend the first on-site course of the project. The laboratory experience gained in the field of new in vitro immunotoxicological tests will enrich scientific development works on exposure to immunotoxic compounds.