Cass Nordmann - Dr. Sandra Demaria - Week 7

 Last Friday, after I had posted my blog for the week, I spoke to Dr. Demaria about brining some organoids to Ithaca. She’s definitely interested in what our more mechanics-focused lab can do with patient-derived cells. We’ll have to work out some of the details, likely after the immersion term is over, but I’m really hoping to continue working with her lab. I feel so fortunate to have found a clinical mentor who fits my research interests so well and I would love for our labs to continue to collaborate in the long-term. One thing that I’ll have to work on if we do try to use organoids in Ithaca is figuring out how we’ll culture them. The experiment I’m doing right now is sort of an early exploration on whether the methods we use with tumor spheroids are compatible with patient-derived organoids.

I was finally able to do the experiment I’ve been trying to get to for weeks on Wednesday. The collagen I ordered still hasn’t arrived in the mail and I can’t conduct the experiment I had planned without it. But finally, Faith was able to ask her clinical mentor Dr. Spector for some collagen on my behalf and I met one of his lab’s postdocs on Tuesday to retrieve it. It’s thanks to them that I’ll have something akin to results to show when I present this coming Tuesday.

There were a lot of issues that came up when I made the collagen gels, so I don’t have too much confidence in the results to be indicative of how these cells behave in specific collagen and cell concentrations. The collagen that the Spector lab uses has a stock concentration of 15 mg/mL, while I’m used to using collagen that’s 3-4 mg/mL. The denser collagen is a lot more challenging to work with. It’s so viscous that it can’t be pipetted like I normally would do so I had to use a syringe to draw it up and use the graduations on the tube to measure it. But due to the shape of the conical tube the collagen was in, it was very difficult to get enough collagen without also getting some air in the syringe. Because of this, I can’t be confident that the amounts of collagen I used for each concentration were accurate and the actual concentrations of the gels aren’t what I originally planned. However, just looking at the images I have so far, I can tell that I achieved a gradient of concentration values. Even if I can’t say from these results how the cells behave in 1.5, 3, or 4 mg/mL collagen I can at least get an idea of what they look like at “low”, “medium”, and “high” concentrations.

There were some other issues that came about like having trouble with how large the spatula I had to stir the collagen was, creating air bubbles while mixing, and having trouble pipetting domes into a small well plate, which is something new to me. But ultimately, I was able to plate some gels and I can see cells in the images I have so far. I’ve placed the plate into a device called the IncuCyte, which is a live cell imaging chamber that’s environmentally controlled. The IncuCyte will take an image of each of my wells every 6 hours so I can track cell growth and organoid formation. So far, the cells seem to be doing alright in the collagen so it might be feasible to use it in the future. I’m hoping this can be something that I work on further this Fall.