Research Highlight: Dominic Kleinknecht

By Kurtis Chien-Young

Dominic Kleinknecht

Dominic Kleinknecht

Dominic Kleinknecht is a Masters student researching at the Fraunhofer Center for Manufacturing Innovation at Boston University. He worked with TuftScope while studying in Boston in 2015.

1. Tell me a bit about yourself. What did you think about your time as an exchange student at Tufts? Where are you working now?

My name is Dominic Kleinknecht, I am a second year masters student in Biomedical Technologies, and am currently working on my Master thesis research project at the Fraunhofer Center for Manufacturing Innovation at Boston University. I’m developing a rapid, field-ready diagnostic test for biodefense priority pathogens such as Zika virus. This is the second time I’m in Boston for a thesis project, after I extended my study abroad during my junior year at Tufts University for a summer research project in the Kaplan lab for my Bachelor thesis. There, I supported a PhD student on a tissue engineered, in vitro human cortical brain model.

My time at Tufts was actually a big factor for me to actively look for options to come back to Boston, since my two semesters as a Jumbo were an amazing experience from start to finish! My undergrad program in Germany in Molecular Medicine had a mandatory year abroad integrated into the curriculum, and my home university in Tübingen has a long exchange history and awesome exchange program with Tufts. I wanted to study abroad in the US, and more specifically in Boston, so Tufts checked all the boxes – and in Fall 2015 I found myself arriving on the beautiful Tufts campus and picked up my keys at the Police Station for my dorm room in Lewis, ready to take on my exchange year! Another cool thing of my exchange year was that I did not have to fulfill any credit requirements for my German program - I could take whatever class I wanted to. I took full advantage of that and thus got to know many amazing people, all from different majors and minors, ranging from Biomedical Engineering to Computer Science, IR, and Psychology. It was amazing, and a very formative year for me that made me fall in love with Boston. And unsurprisingly, I am back again for more. ^^

2. You were recently credited as a co-author on a paper published in ACS Biomaterials Science & Engineering. Could you tell me about this work, as well as how you personally contributed to it?

Yes, the paper was first authored by the PhD student I joined in summer 2016. He was working on developing a tissue engineered human cortical brain model in a petri dish. Therefore, he prepared silk polymer scaffolds that looked like little donuts, in which he wanted to culture human induced pluripotent stem cells. These cells would then be differentiated into neurons or other neuronal cell types to, eventually, mimic a human neural network in vitro. The research was foundational in nature but in the long run, having a tissue model for human brain tissue could be used as a drug testing platform, or a research platform for e.g. traumatic brain injury with more translatable results than animal models. I remember that when I joined the project, it was a 50:50 chance that cells would survive or die, and he didn’t know why. We figured it would be helpful to determine the best possible baseline conditions for undifferentiated stem cell survival in the scaffolds, before differentiating them into neurons. My project over the summer was thus culturing human stem cells, preparing the silk polymer scaffolds to provide the cells a space for spatial attachment, and seeding the stem cells into the scaffolds, maintaining the seeded cells in 3D tissue culture, and running viability tests on them. The experiments I ran were seeding the cells into the scaffolds at defined concentrations, maintaining them for 5 days, and then running experiments on cell viability. After 5 days, my supervisor took over the stem cell-laden scaffolds and started the differentiation process. By the time I left and had finished my project, we had nailed down the optimal seeding and maintenance conditions for the first 5 days to yield densely stem cell-laden 3D scaffolds with viable cells. From there on, he optimized the neuronal differentiation in 3D, kept the growing neural network alive for 6 months in culture, and ran experiments on neural action potential firing patterns in the “little brains,” which there were! The model even responded to physical stress – the firing pattern changed when weights were dropped onto the neural network, just like a mini-concussion! All in all, a very cool project with many promising implications for further research and the future!

3. Why is your research important? How do you imagine the 3D tissue models will be used?

I reckon the main application for the near future is going to be further research. The model is impressive on its own already but could obviously be improved in many ways. We are far from replicating a brain in a petri dish; there is no vascularization, many cell types are missing, and it’s only a couple thousand neurons compared to the millions of neurons in an actual brain. But that’s why it’s a tissue model, after all. It could also be very well used for better drug research or even foundational research into traumatic brain injury. Animal models are the norm, but only a fraction of drugs that show promising results in lab animals make it through the approval process or show any effects in human pilot studies to begin with. Testing drugs on human tissues could be an important building block in expediting active compound research and could also help understanding human pathologies and conditions better. Why study Parkinson’s or Alzheimer’s in rats when you could grow a neural network derived from Parkinson or Alzheimer patients that already come with the exact pathological genetic or enzymatic changes we see in humans? For these points, the tissue models I helped develop could certainly be a helpful and powerful tool.

4. What are some of the problems you faced when doing research? How did you go about solving them?

Research is, at its core, daily problem solving, and I had quite a few in my project. The stem cells kept dying for no reasons, then survived in all seeding concentrations the next experiment, which was baffling. It was the first research project I was part of where I basically had free hand over what I do, and how I do it, so running into issues meant it was on me to figure it out. What really helped was meticulously going over the routine that was performed to set up the experiment. Were the stem cells contaminated? Were the tips not sterile? Was the media expired? Questioning everything was my number one strategy for spotting potential sources of error. I also applied the rubber duck approach to my experiment planning, a concept I was introduced to in COMP11 during my Spring semester. I would talk out my planned experiments, either to myself or to someone in the lab, to have a second opinion on potential issues. And the last thing is not shying away from asking people. Asking never hurts, and my experience has been that scientists are very helpful and encouraging.

5. Do you have any advice for undergraduates who may be interested in your field of study? Is there anything you wish someone had told you before you started working?

Don’t think you need to come in with a ton of knowledge or skills. Science is often just giving it a shot and seeing what’s happening. How did I determine my initial seeding concentrations I tested? I checked some paper, found very different concentrations, tried to adjust it to the size and volume of the scaffold, and ballparked it, basically. Science is an iterative process, which means it is time-consuming, and can be very, very unforgiving, especially if nothing seems to work. But, that’s what I had to learn the hard way in my first couple weeks, even that is a scientific finding, and one piece in the big puzzle that you want to solve. I’ll be honest, science is not for everyone, but if you have a passion and want to gain a deep understanding of what you are researching, you’ll have a great and rewarding time! Personally, looking back, it had helped me a lot that I tried myself out in many different disciplines of Biomedical Research over the course of my academic career through lab courses, internships, and side jobs. I’d recommend to just try it out, get your feet wet in different fields, do a little side project, or join a lab over the summer, and maybe you’ll also catch the science bug. The feeling of finally figuring out why your experiments were not working, or your experiments coming out the way you want them to are hard to beat. 😉

6. What kind of projects are you working on now, if you are allowed to talk about it?

As said, I am currently working on my Master thesis project where I am developing a rapid and field-ready diagnostic test for Zika virus, one biodefense priority pathogen. The end game is a diagnostic device, and we’re very focused on rapid manufacturing and low costs, which guides most of the design and assay development decisions. Unfortunately, I signed a confidentiality agreement concerning the details of my work, but what I can say is that it is an awesome project and that I highly enjoy my day-to-day research routine! The pieces are coming together, and the main assay is already working under lab conditions – next steps will focus on making it more robust to environmental conditions and contaminations and come up with an initial design for the device. Outside my thesis project I also work on a biotech startup as an affiliate, network my way around Boston, and am currently applying for PhD programs. Also, what a surprise, in Boston. 😉