Week 6: Ben

To start of Week 6 of Immersion term, I continued my time in the bariatric clinic. I observed three hernia repairs and one gastric sleeve. The gastric sleeve surgery was pretty standard, but the hernia repairs were different in severity and location, and it was interesting to see the decisions that were being made in regards to how to approach the surgery and prevent its recurrence. 

The following days were spent practicing the islet isolation protocol from mice. I was given 5 practice mice that were euthanized by the graduate student I am working with. The whole process entails cutting to the mice abdomen, identifying the common bile duct, clamping the area where the common bile duct meets the small intestine, and inserting a needle to the duct and injecting in collagenase to digest the pancreas. The biggest challenges were identifying the area in which the duct meets the small intestine, and inserting the needle to the common bile duct in a way in which the collagenase actually flows into the pancreas. Over time and more repetition, I was improving and perfusing the pancreas more and more. Overall, the practice runs were successful.

The next day, I did the protocol on WT mice donated to me by the Alonso lab. With these mice, I intended to isolate islets, disassociate them, and reaggregate them with different amounts of HUVECs and run glucose stimulated insulin secretion (GSIS) to compare insulin secretion capacity between the groups. I was able to successfully isolate the islets from the two WT mice I was given, so I feel very confident in doing the protocol, and feel like I can definitely bring this procedure back to Ithaca so I can perform it by myself. After, I reaggregated the islets in a 0:100, 10:100, 15:100, and 20:100 HUVEC to islet ratio in a V-bottom 96-well plate. I am going to culture these aggregates over the weekend and run GSIS early next week, collect the media, and run an ELISA to compare the insulin secretion response.