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Maya Sim, Weeks 2-3


     Due to some miscommunication and scheduling problems, I got my UCSF ID badge at the start of my second week. Now, I could go to the bathroom without being locked out of the lab and finally start my online training. I successfully completed the training within a day since Nino (my mentor) had to go to the hospital to retrieve thyroid glands and a few other organs from a human donor.

     These samples were frozen and stored in the cold room, which is what lab members call the giant walk-in fridge. The next day, we thawed the thyroid cells and placenta cells to sort them using the fluorescence-activated cell sorting (FACS) machine. The steps required to use this machine is quite long, but I was able to master it after preparing over a hundred samples and repeating the protocol about five times. First, the cells need to be transferred to well plates and centrifuged for five minutes at 1200rpm. Then, I get rid of the supernatant with a sharp throwing motion and resuspend the cells using FACS buffer. Antibodies for CD34 and CD45 are added to each well, then they are incubated in the cold room for 30 minutes. During this waiting period, I have to label FACS tubes for all of the samples and set up the program on the computer. After incubation, the cells are centrifuged once again, the supernatant is discarded, and cells are resuspended. Then, they are transferred to individual FACS tube for analysis. It does not sound like a lot of steps when written like this, but considering there are 96 wells in a plate, I spent quite a long time pipetting.  

     At the start of week 3, Nino and I went to the animal center for the next step of the transplantation. We first anesthetized the 5 mice that underwent surgery to insert the empty encapsulation devices two weeks ago. Then, Nino made a small cut right beneath the previous wound and looked for the catheter. Once he located it, he cut open the film of the device to reveal the end of the catheter. After making sure the device is stable and the catheter is properly aligned, I infused the mouse islets with a micropipette. Then, I sealed the device by cauterizing and stapled the incision. For one particular mouse, we had trouble isolating and stabilizing the catheter. Another mouse kept falling out of anesthesia and moved around during the procedure, so we had to increase the dose of isoflurane. Unfortunately, the last mouse to undergo the cell transplantation failed to wake up from anesthesia, which Nino thought it was because of the increased dose. To check if the cells were infused properly, we took the remaining 4 mice to the imaging room. With the In Vivo Imaging System (IVIS), we confirmed that the beta cells were located within the device.

     For the next couple of days, we went to check their blood glucose levels. Nino was in charge of snipping their tails for a blood sample and I was in charge of using the blood glucose meter and writing down the data. While the normal blood glucose range for mice is 90-120 mg/dL, these mice were around 190 – 230 mg/dL at first and it gradually decreased to around 120 – 140 mg/dL. So far, the results are promising! Along with the bioluminescent images from IVIS, we are planning on keeping track of these mice for a few more weeks to see if diabetes is reversed. Meanwhile, I am anticipating to test a newly designed encapsulation device with a solid ring for structural support as soon as we receive the devices.
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