Submitted by Amanda Wu
I’m excited to say that I’m finishing up my third of seven weeks at Cedars-Sinai! Time flies by fast, doesn’t it?
Since the previous installment of my blog update, much has occurred in regards to the progression of my research project, the focal point of which is based largely on Huntington’s disease and stem cells. As you may or may not be aware of, Huntington’s disease is a genetic disorder that causes the progressive deterioration of nerve cells within the brain, inhibiting and resulting in the loss of cognitive, mental, and motor capabilities.
My research during this summer emphasizes primarily on the differentiation of induced pluripotent stem cells into medium spiny neurons, cells within the striatum that are most directly affected in individuals afflicted with Huntington’s disease. As such, my research as it pertains to Huntington’s disease entails how varying durations (1 wk., 2 wks., 3 wks., and 4 wks.) of the application of doxycycline to induce gene expression to induced pluripotent stem cells affect the quantitative yield of neural progenitors. In simpler terms, I am attempting to discern the duration of the application of doxycycline to induce gene expression for the optimal yield of neural progenitors derived from the induced pluripotent stem cells.
In order to discern the optimal duration of doxycycline application, much of my efforts within the past two weeks have been largely focused on counting cells. Specifically, I’ve been counting fluorescent markers indicative of neural progenitors, cells that are likely to have a tendency to differentiate into neurons, as well as neuronal markers in microscopically imaged cells treated with doxycycline for varying durations (1 wk., 2 wks., 3 wks., and 4 wks.) through the utilization of a program known as ImageJ.
In addition to the attainment and analysis of data through cell counts, I have also observed my mentor and her performance of tissue culture before performing my own ICC. If I’m being honest, when I first heard about ICC, the history nerd in me initially registered the acronym as the Interstate Commerce Commission from the Cleveland administration. However, that meaning is obviously not applicable to ICC in this case. Rather, ICC signifies immunocytochemistry. In order to attain the fluorescent tags for imaging and counting the cell markers, the cells treated with doxycycline were first fixed using paraformaldehyde and then stained with fluorescent primary and secondary antibodies that bind to the markers.
One takeaway I’ve captured from ICC is that it’s okay to break a few eggs, or rather a few cover slips, to make an omelet. All jokes aside, I’m eager to observe further progression of my research.
Hope to keep you updated!