Week 7: Ben

 This week, I continued to work on my islet reaggregation experiment, where I recombine disassociated islets with different amounts of human umbilical vein endothelial cells (HUVECs). Specifically, I combined 1 IEQ (islet equivalent, approx. 1000 cells) with 0, 100, 150, or 200 HUVECs as literature reports that islets are comprised of about 10-20% of endothelial cells in number. I put these cell suspensions in a V-bottom 96-well plate where they will reaggregate in the bottom. This was set up last week, and I let the spheroid formation occur over the weekend. However, when I came into lab on Monday to check on them, I found that they did not form into spheroids properly. Reflecting on it, I realized that I forgot a coating step in which I incubate the plate with a ultra-low adhesion solution that prevents cells from adhering to the well, which is the reason that the spheroid did not form properly. Either way, I moved forward with my plan to run glucose stimulated insulin secretion (GSIS), as this assay will be used to test my original hypothesis: that more blood vessels leads to increased insulin secretion. Even though spheroids did not form, there is value to running GSIS on what is essentially a 2D co-culture, as I can check if there are any secretome interactions between the two versus matrix interactions which was what I was originally aiming for. With this, I ran GSIS, which entails a 1hr starvation period, a 30 min low glucose stimulation period (3mM), a 30 min high glucose stimulation period (20mM), and a lysis period which released all insulin in the intracellular granules. In the low, high, and lysis steps, I collected the media for analysis on an insulin ELISA to measure the total insulin secreted in these steps. Comparing insulin secretion in these steps between by different HUVEC density groups will give me hints in how islets and endothelial cells interact. Hopefully, I will have this data before my presentation next Tuesday.

In addition, I followed my clinician Dr. Alonso to her clinic. This clinic was interesting because there were a lot of cases in which patients had type 1 diabetes but also were going through pregnancies. It was really interesting to see how diabetes management and care is different and so much involved and crucial in these cases, as extra precaution is needed to manage blood glucose. What was fascinating about these cases was learning the biology from Dr. Alonso about how the pancreatic islets adapt to increasing nutrient uptake and metabolism to help the fetus grow.

Moving towards next week, I will wrap up my lab experiments, attend one more clinic, and get ready to head back to Ithaca. This summer experience has been eventful and eye-opening, and I definitely have many ideas to apply to my thesis work. I cannot believe Immersion is almost over!