Week 4: Eleana

This week was very eventful. Over the weekend, I made some biocompatible hydrogels from Alginate in Ithaca and carefully transferred them back to Weil. On Monday, I then functionalized the hydrogels with two antibodies anti-CD3 and anti-CD28. These antibodies are required for T-cell activation and proliferation. Tuesday, we started with a T-cell extraction from the spleen and lymph node extraction from mice. We then isolated our cells, purified them to only have T-cells that expressed CD8+ and plated them on hydrogels. The activation takes three days, during this time they are seated on the gel and on an oscillating shaker to provide mechanical forces during activation which T-cells prefer. During this waiting period, I passaged SB28 glioblastoma cells and let them grow in culture. I transferred them to a plate that would be able to fit within the irradiator. We also had the ability to go to a conference where two people presented their work on prostate cancer and cancer organoids with immunotherapy. It was a very insightful conference and some work was heavily genetic focused while others were focused on the treatment and immunotherapy side. On Thursday and Friday we went back to pathology. During this time in pathology, we saw many patient cases with DCIS (ductal carcinoma in situ) and LCIS (lobular carcinoma in situ). In many patients it was very difficult to tell the origin, that is why pathologists also perform an E-cadherin stain to differentiate the two. There was a classical example of LCIS which looked similar to what I was studying in the textbook and I was happy that I was able to identify the type. Throughout the weeks I have been studying the histologic and pathological differences of the breast cancer subtypes those that are benign and and malignant. I have noticed in an improvement the more I look at the slides to identify atypical features within the tissue. Friday we began irradiation of gliblastoma cells with co-culture of T-cells which was an all day process. We wanted to irradiate the SB28 cells and co-culture them with T-cells to see if the gels were able to prevent T-cell exhaustion as compared to no gels. We will confirm these results next week after co-culture is complete and with flow cytometry.