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| Bowling with friends |
On the first day of week 3, Shawn and I changed media and did von Frey testing on the male mice. As usual, their pain threshold was higher than the female's and they were less fidgety. The following day, we sacrificed 10 more mice: 5 males and 5 females. We plated the bone marrow cells as we typically do and continued to change the media for all of the batches of cells.
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| Shawn removing bone marrow using a syringe and tweezer |
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| The hood where we change media and plate cells |
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| Additional image of female BMSCs plated on 6/14/17 |
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| Image taken on 6/30/17 of female BMSCs plated on 6/14/17 |
On the Wednesday of week 3, we made new media because we were running low. We put two antibiotics, penicillin and streptomycin, and fetal bovine serum (FBS) in the media. Though I knew why we put the antibiotics in the media, I did not know why we put FBS in. As it turns out, the FBS provides proteins that are necessary for BMSC growth. In addition, we prepared and distributed the chemotherapy drug, paclitaxel, into smaller amounts so it could be ready for injection. Following this, we changed the media of all the cells and bought supplies, like filters and pipettes, for the lab. Because we had just plated new cells the day before, we also had to wash those specific cells with PBS to clear out the unwanted cells. Thursday, we sacrificed 10 more mice and plated their cells with media. On Friday, I finally got to meet my PI, Dr. Ru-Rong Ji, who had just returned from China. We had a lab meeting and it was rather long because Dr. Ji had to get caught up on the details of everyone's research. I learned a little bit about other people's research and at the end, Dr. Ji gave us Swiss chocolate that he got from Switzerland. Once again, we changed the media and gave PBS to the plates from the 29th.
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| Plate of BMSCs |
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| Magnified image of BMSCs |
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| Female BMSCs from 6/27/17 |
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| Female BMSCs from 6/27/17 |
On Tuesday, Shawn, Deja, and I went bowling at an alley close by. After work on Friday, we went to the Duke Lemur Centre and saw a variety of cute and loud lemurs. Saturday, we went to the Duke Gardens before Deja's mother and sister arrived. The botanical garden was very pretty and had a wide array of plants. As usual, we had to go to Elon to pick up more tacos at our favourite place. This time, Shawn tagged along and we brought some back for Deja's dad and sister. Sunday afternoon, Deja's family and I met one of my best friends at the North Carolina Museum of Natural Sciences. This was the second time I got to see my friend and her baby who live right around the corner in Raleigh. Surprisingly, there were labs in the museums that are just like ours in EXP- pipettes, centrifuges, wells, and everything else. At night, Deja's sister, Deja, and I went go karting and shopping at the mall, and to make the night extra special, we finished it off with a trip to Only Burger for some delicious food.
During week 4, I was not as optimistic as I had been before. Though the week started off right, I was fueled with anxiety the days following July 4th. On Monday, we had another lab meeting, but this time, it was brief. We also replaced the media in the wells and did behaviour testing on both males and females. This testing was really important because it would serve as the baseline for our experiment. Following lunch and a quick run to Starbucks, Yul injected paclitaxel into 10 male and female mice in their abdomen, carefully avoiding the digestive organs. Because we were running low on anesthesia, only half of the mice were injected under anesthesia. That night, Kaitlin, my friend from Raleigh, picked me up and we had a sleepover. The best part about it was the ice cream at a place called Sunni Skies in Angier- totally worth the 20 minute drive from her house. On Tuesday, I spent my day off with Kaitlin, her mom, and baby at Dave and Busters and her house. We finished off the night the only way we knew best: more ice cream from Sunni Skies. Though I enjoyed my holiday, the cells did not. Our cultures seemed to be doing very well before the holiday, but as soon as we came in the next morning, we noticed that a portion of our cells were missing; physical plates were not missing from the incubator, but cells inside the plates were missing. Ever since the beginning, we had not had any issues- no contamination, no mishaps, nothing. Now, all of a sudden, our plates seemed to be see-through. As usual, Shawn and I were jokingly blaming each other for the disappearing cells even though it was no laughing matter. Stunned, Yul could not figure out what was wrong; our techniques have been consistent and sterile. In addition, it was obvious that there wasn't a bacterial outbreak because the wells were not cloudy. One of the other members of the lab mentioned that the loss of cells may be due to a fungus, so now it was time to make a new media; this time, an antifungal would be added. Throughout the rest of the week, we did behaviour testing on the mice with paclitaxel and injected them again on day 3 and 5 (we later injected them again on day 7). We sacrificed 10 more mice on Friday so there could be a fresh batch of cells with this new and improved media. We still kept most of the other plates of cells that looked as if they could be promising with the new media. Now, Yul decided that he should change the media for the new culture to rule out the possibility that Shawn or I made a mistake. As it turns out, the disappearance was not me or Shawn's fault because the wells we replenished with new media started growing back. Now, the cells are thriving and look as if some are ready to be taken out and injected into mice.
That Friday, one of my aunts came to visit me from Jersey. Because her flight got in at 8 a.m. and I did not have to be in the lab until 1 p.m. I went out to breakfast with her at a place called Monuts. After that, we went to Duke Gardens and walked about 5 miles- fortunately, it wasn't as hot as usual, but it was still pretty gross. After work, we went to a place called Bad Daddy's in Raleigh for dinner with Kaitlin and her family. We had an amazing dinner and my bean burger was outstanding. The next day, my aunt and I went to downtown Raleigh for the day and went mini golfing at a place called Frankie's; I now understand why there are so few places to mini golf in North Carolina- it is way too hot to be mini golfing. After that, we went to Kaitlin's house for dinner and got our nails done in town. Like any other day in the Raleigh vicinity, we ended it with a quick stop at Sunni Skies for ice cream. On Sunday my aunt and I went on a shopping spree at the mall and got wings (or vegan wings, in my case) at a place called Heavenly Buffaloes- and they were
heavenly. Sadly, my weekend with my aunt had to come to an end and she flew back to Jersey on a 2.5 hour delayed flight.
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| My aunt, Kaitlin, her mom, baby, and me at Bad Daddy's |
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