Kabir Grewal, UPitt Lab, End of Week 4



I ended my last blog post with writing about when I injected TMSC’s and CSSC’s into  well plates with  the extracellular matrix of bovine corneal endothelial cells (BCE cells) , which brings me to the work I completed last Thursday and Friday.

Last Thursday was a busy day for me.  After arriving at the lab at the usual time of 9:30, I checked the growth of the newly injected stem cells mixed with the extracellular matrix of lysed BCE cells. As expected, the stem cells had not attached to the bottom of the well plates, and were free floating in the DF10 growth medium. Today, my task was to make a new growth medium for these cells, as well as create the variables for the experiment.  The new growth medium I made comprised of a glucose-rich base, fetal bovine serum (FBS), as well as ROCK inhibitor (short for Rho Kinase inhibitor). The purpose of this inhibitor is to prevent excessive apoptosis among the cells, and promote the cells to grow and retain their structure.  The variable for this experiment is including aqueous humor to half the cells, to see if the nutrients in the humor could potentially induce the stem cells to differentiate into BCE cells more efficiently.  So, in half of the 8 wells, I included aqueous humor.  Then, every hour, I monitored the cells to make sure they were responding well to the new medium, which they were. 

Where I spent most of my time this week- working with these microscopes to observe the stem cells


On Friday, Dr. Peretz came to visit me! It was fun showing her around the Du lab as well as some of the work I have been doing.  After this, we enjoyed a delicious lunch at a restaurant called Burgatory, which is basically the Pittsburgh version of Shake Shack.  It was great to hear about the work some of the other EXP members are doing this summer. After lunch, I proceeded to check on the cells again. Specifically, I was looking to see if any of the stem cells began to attach to the bottom of the wells. For the most part, they had not, so I put them back in the incubator to let them grow more.  I left work earlier than usual today at around 4:00, and went back home to my Aunt and Uncle’s house to babysit my 8 and 10 year old cousins.


Going to work every day is definitely getting more and more exciting now that I am able to perform experiments independently.  My experiment is very different from those of my lab mates, so it is neat that I get to explain to them what I am doing.  I find myself more and more engrossed in the work since I spend the time in the lab either running my experiment, or reading up more on certain aspects and discussing them with Dr. Du.  It was certainly one of the best weeks I have had so far.  

Comments

Post a Comment

Popular posts from this blog

Stephanie Wu | Week 3 | More Slicing and Mounting

Image
06.19.17 - 06.25.17 For the majority of this week I spent my time slicing and mounting the brains Felicia had previously removed from some mice via perfusion after inserting the injections she was working with. The main type of mouse I have worked with are the rAAV2 Delta-Cre mice (which are the controls) and the rAAV2 GFP-Cre mice (which are the experimentals), which are then divided into hetero and homo. From there, Felicia targeted specific regions of the brain, mostly injecting into areas such as the nucleus accumbens (NAc) and the dorsal striatum (DS). She injected both a control virus as well as multiple experimental viruses. By slicing the brains we would eventually be able to image them on the microscope and see the effects of the injection on the synapses of the brains and the different afferent sites as well. (some brain slices (N1C. 87)) (more slices) (some mounted slides of N1C. 125 & 126 brain slices) (what my slide box currently looks like) Bes...
Read more

Emily Guo, Week 7, Hanging Up the Lab Coat

Image
Some good news: the plasmids Anna and I made a few weeks ago were sent off for sequencing, and they came back with the correct sequence! The plasmids can now be used in the next round of electroporation.  We are currently still troubleshooting the bisulfite conversion process. I ran a PCR for the DNA samples (non-converted) that we were using on Monday with two sets of primers that we knew would work, and we did indeed have bands of DNA show up on the gel afterwards. Therefore, the quality of the DNA that we were converting wasn’t the problem. I found a bisulfite conversion guide by Zymo Research – which deals a lot in epigenetic research in addition to providing biology research tools for DNA and RNA research and analysis – that provided some tips that we could try out. One was a way to analyze the DNA after bisulfite conversion, which we didn’t do before, by running the gel with the bisulfite-converted product. However, because the DNA becomes so damaged through the conversi...
Read more

Andrew Mah, Entry #1, Totally Unexpected

Image
My first two weeks at Khalizov’s lab just came to an end. The transition has been somewhat rough since there were many unexpected parts of the lab I was not prepared for. But, I am trying my best to get myself back on track. Anyways, before I get to the actual lab experience, my daily schedule at NJIT goes from sometime between 9:30 and 10 in the morning to 4:30 in the afternoon. My lab is really flexible about times, so this has been great for me. Commuting from home takes about 30 to 40 min since the traffic to Newark is always packed. I have tried many different ways to get to NJIT: train, my mom/dad dropping me off, and Uber. Now, going back to the lab experience, I thought I would be prepared for the lab since I visited the lab back in February and also got the lab safety training out of the way early. However, the lab turned out to be a more of an applied physics/engineering lab than an environmental chemistry lab. The lab is very small in te...
Read more