Team 4

Effects of Exposure to Sanicle Essential Oil on the Cellular Proliferation of Squamous Skin Cell Carcinoma
Team Members:

  • Aaron: I’m a driven and involved student, and I’ve recently become very interested in biology. In middle school, I became interested in biology and learned about the Rock Canyon biotechnology program, and I went to Rock Canyon with the goal of getting into the research program. In the future, I hope to go into the field of biotechnology.
  • Carter:The best way to put this is that I basically live for science, because if science lets you live, then why not live for science? My interests in science range from theoretical particle physics, to chemistry, to set in stone clinical medicine, however, physics and biomedical research have always topped the other subjects for me. With aspirations in science like becoming a trauma surgeon, or doing biomedical research, I am super excited to take the Biotechnology Research course this year to learn useful lab skills and conduct real world research on one of the world’s most prominent diseases, cancer. Outside of my ambitions to change the world in science, I love the outdoors and have a great passion for cross country running, swimming and triathlon!
  • Griffin: I am a very motivated student athlete who enjoys getting involved in as many science classes and clubs/sports as possible including Honors Chemistry, AP Chemistry, AP Environmental Science, Biotech 1, Biotech 2, Honors Biology, baseball, Chemistry and Science Olympiad, etc. I wish to use the knowledge and skills I develop though these classes and extracurricular activities to benefit me in my future in pursuing a position in the medical field. I took this class to dip my toes into the field of research and to gain the experiences and relationships that are unique to this class.

Project Goal

As of  Date 9/10/18

It's Finally Working!

posted Feb 8, 2019, 7:57 PM by Student Aaron Sonin   [ updated Feb 8, 2019, 9:13 PM by Student Carter Brand ]

Hello, everyone! If you have been keeping up with our blog post, you would know that we haven’t had the best relationship with science lately. Luckily, it seems like the tides have finally turned. To celebrate, and in order to insure the best experience while reading this blog post, please play the song below.

YouTube Video


We spent the end of last semester and all of winter break poring over our procedures, trying to figure out what we could be doing wrong. When we got back from break, we still didn’t know exactly what was going on. But thanks to Ms. Emily Duncan, our mentor, we decided to check the incubator. Since the incubator is pretty old, it isn’t in the best shape and therefore, the CO2 sensor needed to be calibrated. We calibrated the incubator and restarted it, allowing it to measure the CO2 and temperature levels accurately. Once we calibrated it, we started growing some NIH3T3 cells (the noncancerous line) to make sure that everything was working properly.

And lo and behold, everything worked perfectly! It turns out, the CO2 incubator was the problem all along. Because the incubator wasn’t calibrated properly, the CO2 levels, temperature, and humidity was reported by the incubator as being at the proper levels, while in actuality, they were not. And because the cells require specific conditions to survive in cell culture, they were not growing properly. We don’t know what caused it to become uncalibrated in the first place, but it’s most likely that it’s just due to the age of the incubator and the sensors being worn out over time.

So, with everything working properly, it was finally time to start trials again. We started with NIH3T3 cells (our noncancerous line). Just like before, we treated the plates with either nothing (the control group), glycerol (a control for an ingredient contained in sanicle oil), or sanicle essential oil. In addition, the glycerol and sanicle plates were treated with DMSO, which allows the cells to absorb the oil. So far, we’ve done three trials of NIH3T3 cells. However, the data from the first one cannot be used, because an unexpected snow day prevented us from counting the cells, creating a hole in our data.

So far, we’ve noticed that the NIH3T3 cultures treated with sanicle proliferate, or grow, at a significantly slower rate than the control cultures treated with either glycerol or nothing. Interestingly, there were no dead cells in the cultures treated with sanicle meaning sanicle oil does not kill the cells, but instead slows their rate of proliferation. In addition, the cultures treated with glycerol are proliferating at a similar rate to the control plates treated with nothing. This means that neither the glycerol nor the DMSO affect the proliferation rate of the NIH3T3 cells, meaning that whatever is affecting the growth rate of the cells is a component of sanicle itself. We haven’t tested A223 cells (the cancerous line) yet, so we don’t know if the sanicle affects the A223 and NIH3T3 cell lines differently. We will be performing the A223 trials over the next few weeks!

We hope to wrap up our research and run statistics in the coming weeks, and we hope to get interesting results. Stay tuned!

Trials and tribulations

posted Dec 14, 2018, 11:04 PM by Student Aaron Sonin

Hello everyone! It’s that time of the year again, the holiday season! And for most people, that means bundling up, getting a warm mug of hot cocoa, and maybe even building a snowman in the yard. But for biotech students, the holiday season means working hard to finish our research before winter break starts.

If you recall from last time, we ran into a bit of trouble when starting our trials. After thanksgiving break, we got right back to work and started culturing cells for our next trials. Since we were running behind on time for trials, we plated both A223 and NIH3T3 cells to be cultured and split on Friday. But for some reason, our A223 cells did not grow, so we contacted Emily Duncan (Our mentor at CU Anschutz) and picked up more A223 cells the following week. On Friday, we split our NIH3T3 cell culture into 15 plates for our trial. Besides that, there’s nothing particularly exciting that happened that week. So let’s get to the exciting stuff: trial 2!

For trial 2, we reduced the amounts we used to treat the cells as we said in our previous blog post. We started on Monday, December 3rd, by treating our experimental groups with the Sanicle essential oil, and glycerol (an ingredient that comes in the sanicle oil that we’re controlling for). Throughout the week, we counted and treated the cells every day. However, about halfway through the week, we noticed something strange. The cells’ growth was stagnant, even in the untreated control plates. We tried giving them some hot cocoa and a scarf and put them in front of the fireplace to warm up, but they still didn’t grow. (Okay, maybe we didn’t do that. We actually just washed the plates with Phosphate buffered saline when changing the media to get any potential contaminants off of the plates.) But, we had plated A223s earlier in the week for a trial this week, so we continued on thinking that it may have just been a fluke. We split the A223s into 15 plates on friday to be used for the next trial.

For trial 3, we used the same amounts of treatment last time since it most likely wasn’t the treatment killing the cells, since the untreated plates didn’t grow either. We observed, counted, and counted the cells every single day, just like the last two trials. However, the cells didn’t grow in this trial either. There were lots of dead cells in the plates, even the untreated control ones. We still haven’t figured out exactly why this has been happening, but over winter break we’re going to closely examine all of our methods and see where things might be going wrong. After break, we’re going to continue doing our research and (hopefully) get all of our trials done.

Many Things to Be Thankful For

posted Nov 16, 2018, 6:21 PM by Student Griffin Hayrynen

Hello! As thanksgiving approaches, we thought it would be fitting to count some of the things in this project that we are thankful for.

The first thing we’re thankful for: we narrowly avoided catastrophe! About a week after our last blog, everything was going fine. But suddenly, some suspicious looking substance showed up in some of our cell cultures, which can be seen in the picture. We didn’t know what it was, but after talking with our mentor from University of Colorado Anschutz, Emily Duncan, and our mentors from the previous year, Madison Dittmer and Kylie Hutchison, we came to the conclusion that it was most likely some kind of contamination. Now, this doesn’t really seem like something to be thankful for, does it? But, we narrowed down the source of contamination to the outside environment, and luckily our media wasn’t contaminated. This meant that we only had to wipe down the room, and we started again the next day with new cultures. If our media had been contaminated, we would have needed to make more media, and possibly order new ingredients. It could have been a lot worse!

The next thing we’re thankful for: our A223s are growing now! If you remember from our last blog post, we were having trouble growing our A223 cell cultures. They were growing way faster than we anticipated and were becoming over confluent after a day or two. Now, we’ve figured out just the right amount of A223s to plate in order to get a healthy culture going!

The last thing we’re thankful for: we’ve finally started trials! …Kind of. On Friday last week, we split our noncancerous NIH3T3 cell line into 15 plates for our experimental trials with treatments. Over the past week, we’ve been treating these plates with sanicle oil, along with our controls for glycerin (a chemical that the oil contains as a preservative), DMSO (a chemical which allows the cells to absorb the oil), and controls with no treatment. However, we quickly realized that the dosage of sanicle oil that we were using was killing the cells we were treating it with, which can be seen in the picture. We couldn’t test dosage in pre-trials, since that would allow us to intentionally affect the results of our trials. However, after break, we’ll be doubling up and treating both A223 and NIH3T3 with lower dosages so we can accurately observe the effects of the oil on both cell lines.

Hope everyone has a great thanksgiving, and don’t forget to eat lots of turkey!

Spooky Cells

posted Oct 26, 2018, 3:23 PM by Student Aaron Sonin   [ updated Oct 26, 2018, 8:16 PM by Student Griffin Hayrynen ]

Hello everyone! Before we get this blog post started, please play the music below while reading this blog post for maximum effect:

To kick off the spooky month of October, we had a spooky encounter in our cell culture room with our CO2 incubator! On Friday the 13th, (it was actually the 5th, but pretend it was the 13th) we walked into the cell culture room, to find that the CO2 incubator was disconnected from the incubator by a ghost! We suspected that the ghost still inhabited the CO2 incubator because it was trying to communicate with us by beeping constantly, loudly, and annoying us beyond belief and took the spirit out of us. So we called in the ghostbusters (Airgas, the CO2 company) to figure out what happened and to replace our CO2 tank, but they weren’t available to come in until the middle of fall break. So, we tried to figure out exactly what the ghosts had done to our incubator ourselves. We figured out that the CO2 tank wasn’t installed correctly the first time, and a ghost came in and loosened the connecting tube enough for it to come off of the incubator. During fall break, we set up the CO2 incubator again and plated cells from both cell lines (A223 - Cancerous cell line and NIH3T3 - Noncancerous cell line) without any more paranormal encounters.

However, when we came back from fall break, we took a look at our cells and noticed some spooky behavior. When we plated the cells over fall break, some ghosts laced the media which caused the cells in the noncancerous NIH3T3 plates to grow abnormally fast and become overconfluent (there were more cells than could fit in the plates). Also, some

ghosts must’ve messed with our cancerous A223 culture, because all of the cells on our A223 plates were dead. We were off to a rocky start, but we want the ghosts to win and get their way.

We plated two more vials of each cell line, and we made sure to have proton packs at the ready in case someone tried to attempt any spooky shenanigans. When we checked on our NIH3T3 cells the next day, they were growing perfectly! We’ve kept our NIH3T3 cultures going steadily since then. Sadly, our A223 cultures were still a graveyard. We consulted our mentor and ghost expert, Emily Duncan, for advice, and we figured out that since the A223 cells are cancerous, they grow super fast and they were becoming overconfluent overnight, allowing ghosts to mess with them. There were still a few live cells left in our A223 cultures, so we’ve been nursing them back to health, and things are looking good for them now. We also plated a bit more A223s, and those are doing well too.

We’re almost ready to move on from pretrials to experimental trials, and we can’t wait to get started! Hopefully once October is over, we won’t have to worry about ghosts anymore.

Some pictures of what we've been up to:

Carter preparing dishes for cell culture.

Griffin plating cells into the culture dishes.

Aaron replacing media in cell cultures.

A clump of dead A223 cells. Spooky!

Some healthy cancerous A223 cells

An overconfluent NIH3T3 culture. Those cells look crowded in there!

A healthy NIH3T3 culture.

PS: We almost forgot one more thing. Because ghosts were messing with our cells, we found a few cells that were possessed and grew double, triple, or even quadruple nuclei! (Or maybe instead of ghosts it's a natural occurrence in cell cultures, but we're not sure) Check them out:

Ready... Set... And almost go

posted Oct 5, 2018, 1:01 PM by Student Aaron Sonin   [ updated Oct 5, 2018, 1:22 PM ]

Hello everyone! These first few weeks since the pitch have been slow, but we’ve been itching to get our research started the whole time! After we got accepted, we started preparing to start our pre-trials and the rest of our research. We’ve been busy getting our skills approved, such as plating, splitting, and counting the cells as well as practicing skills we’ll be using during our research. We’ve pretty much gotten that all out of the way, so we’re almost ready to start our research!

This past week or so, we’ve been up to a lot. We got our CO2 tank in, which we had hooked up to our incubator that we’ll be storing our cell cultures in. We also got our liquid nitrogen, which keeps our storage vials of cells at a nice, cool −195.79 degrees celsius.

We also got our cells this Monday, which were donated to us by Emily Duncan (who has also been mentoring us throughout our planning and research) at the University of Colorado Anschutz Medical Campus. We took the whole class period on Monday and road tripped it down there. Emily showed us around the lab she works in, and showed us where everything was.

She gave us our cells in a styrofoam container with dry ice inside to keep them cool. They also come in these neat little cell culture vials:

When we got back to school, the cells were put into a -80 degree celsius freezer, and then the next day we transferred the vials to the liquid nitrogen that we have in our lab.

In order to get our cell culture room ready for the cells, we had to sterilize the whole thing to prevent contamination during our research, and we also started up our CO2 incubator. That involves spraying a lot of isopropyl alcohol everywhere to kill all of the microorganisms that might be hiding in the room. I think spraying down that room took at least twenty years off of our lives.

(Aaron's not the most photogenic person.)

Anyway, over fall break, we’re planning to plate different amounts of cells for our pre-trials to figure out how much we need to use! We want the cells to reach 80% confluency (coverage of the cells on the dish) by the end of the trial, so they don’t start slowing down their own growth and affecting our research.

Until then, this is group 4 (oily cancer) signing out.

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