Through their lens: Andrew Moreno gets inundated with information, and learns about tooth formation

This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.

Andrew Moreno did a stem cell research internship this summer in the laboratory of Jill Helms at Stanford University.

Andrew Moreno in the lab. He submitted this photo to our #CIRMStemCellLab Instagram feed.

Before SIMR, my very limited laboratory experiences left me with the impression that research was the rote adherence to an established process, a mere repetition of the same task with a growing conviction that the outcome will be different if and only if one is more precise with each attempt. This preconceived notion regarding the true nature of research lacked the very thing that made science exciting and worth pursuing: an ardent desire to further what has already been proved in order to better and improve the world. Cliché, I know. Anyway, I decided to apply to SIMR in order to acquire some understanding of what research truly entails, but I accepted my admission to the program with lingering apprehensions and innumerable misgivings. My endgame was to determine whether or not medical research appealed to my interests in the same way that clinical practice as a physician did.

My first day of lab made me wish I hadn’t made the trip to Stanford for the summer. I was first introduced to my mentor and was put at ease on account of how reserved she was. This false security proved to be particularly transitory as the briefing that ensued inundated my sensibilities with talk of PBS, Cre-lox recombination, and Hedgehogs. In a matter of five minutes I was shaken to the core as the project I had elected to take on for the summer was expounded on from every angle. I had mistakenly espoused a self-assured confidence in my educational background and faced the unforgiving reality that I did not and could not know everything. In short, I was lost, confused, and disconcerted all at once.

Of course my perspective was completely skewed and after learning more about the molecular underpinnings of my project, I could better understand the reasons behind what I was doing in the lab. After reviewing the already published data regarding craniofacial development and the role Wnt and Hedgehog signaling have in facial patterning and morphogenesis, I was able to gain direction in the research I was conducting. I came to realize that the deletion of an intraflagellar transport protein, Kif3a, in the primary cilia of neural crest cell derived mesenchyme cells of precursory teeth resulted in an atypical phenotype characterized by an enlarged, invaginated enamel organ. For this reason, we focused on the very early stages of odontogenesis to see what could have engendered the observed patterns. So, we conducted immunostaining for various downstream target genes of the Wnt and Hedgehog pathways to visualize their activity and correlate it with the abnormal phenotypic expression.

In retrospect, I had an enormous learning curve to make up for, but I figure that any new experience comes with one. If I took away anything from my experience in SIMR, it is the realization that there is always something different and exciting to learn in a lab because searching to explain the unknown stimulates one’s imagination. Although I entered this program with an insular and cynical view of research, I find myself looking beyond the scope of what I would have originally thought possible, equipped with a new mindset and a new way of thinking. With this perspicacious outlook, I find myself better prepared for any prospective research opportunity, and as a result, I am so much closer to uncovering what true research is and represents.

Andrew Moreno

Andrew submitted this video about his experience:

Through their lens: Caroline Desler experiences the excitement and disapointment of medical research

This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.

Caroline Desler did a stem cell research internship this summer in the laboratory of Julie Saba at Children’s Hospital Oakland.

Caroline Desler working in the cell culture room. She submitted this photo to our #CIRMStemCellLab Instagram feed.

Exploration, curiosity, failure, and excitement have all marked my research experience at CHORI thus far. These past few weeks, I have experienced what it is like to play a part in a legitimate lab environment—creating hypotheses, thinking “outside-the-box,” learning new (and at first intimidating) procedures, collaborating with other lab members, and oftentimes, falling short in my experimental endeavors. However, I’d have to say that the most enjoyable part of my internship has been working with my mentor, Dr. Aguilar, and getting to know all of the other post-docs and summer students in the Saba lab. Despite their varying origins across the world, these people bring together their incredible knowledge, insight and experience in the research field to make advancements toward a common goal. Seeing the incredible collaboration that takes place in the lab and its power to yield huge levels of success and discovery never fails to amaze and inspire me. This opportunity, made possible by CIRM, has given me a “peek” into what it would be like to be a researcher. Prior to this summer, I viewed research very simplistically—as the boring stuff that takes place in labs without much human interaction. However, I now know that a career in research holds significant importance and value from a variety of perspectives. Research allows us to live healthy lifestyles, free from the threats of many diseases. How else do groundbreaking vaccinations, which have the power to protect and save the lives of thousands of people, develop in the first place? Research explains why things happen. How do we know so much about cholesterol and its role in heart disease? Research serves as the fuel for discovery and increased understanding. No wonder we now have access to our entire human genome! Despite my always wanting to pursue a career as a clinical doctor, I now recognize that all of the clinical work would be largely impossible if it weren’t for the people who devote their lives to research. I look forward to delving further into my scientific interests in high school, college, and beyond—hopefully leading me to a career involving both research and clinical aspects of medicine.

However, that isn’t to say that research isn’t incredibly tough. Already, I’ve had many tastes of the waiting, anticipation, frustration and failure that accompany a career in this field. For example, it wasn’t until my 4th or 5th western blot that I finally got a hang of the technique and after a total of 8-10 hours of procedural work, actually wound up with an x-ray film that looked halfway decent. The sight of my protein bands coming out of the developing machine, nice and straight and even on the film paper, was such a rewarding and magnificent moment. Just this week, my mentor and I went merrily strolling into the cell culture room to take a look at our cell plates, only to find that each one had been completely contaminated by bacteria. Sadly, the bottom of the biohazard trash can was the fated destination for these cells which Ana and I had been cultivating for the past 3 weeks. I can’t even begin to describe the frustration I felt at that moment. However, the recollection that we had just frozen cells of the same cell lines a few days earlier was quite a relief. It taught me that despite the mistakes I make, there’s always a way to move on and make them better…and oftentimes, there are methods to prevent issues like this from happening in the future.

Overall, my internship experience this summer has opened my eyes to the beauty of research, the possibilities of human collaboration, and the importance of failing…for it is often through failure that we best discover new things.

Caroline Desler

Caroline sent us these videos of her experience:

Through their lens: Alfonso Barraza works on a Huntington’s disease therapy, learns the heartache of failed experiments

This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.

Alfonso Barraza did a stem cell research internship this summer in the laboratory of Jan Nolta at the University of California, Davis.

Alfonso Barraza working in the lab. He submitted this photo to our #CIRMStemCellLab Instagram feed.

First of all hello. My name is Alfonso Barraza. I have been working in the Nolta Lab at the UC Davis Institute for Regenerative Medicine. I have pretty much my whole summer been working with the one and only Vector Goddess, also known as Karen Pepper. First day was alright (not really). I mean I got to meet Scary G which was pretty awesome. To be honest I was kind of scared to meet him (don’t tell him that) because he is the UC Davis program director. I really didn’t know what to expect. But I tell you once you get to know him, he is awesome. I also had to go through all of the paperwork and “necessary” stuff for working there. There was a lot less than I thought. THANK GOODNESS!! Going through biosafety training was enough work in itself. Don’t get me wrong the lady who was teaching us everything and telling us what to do when you spill chemicals was really nice. But it’s really hard to make something like biosafety training fun and enjoyable (I almost fell asleep).

The next day was awesome because I finally got to meet her Vectorness. Boy is she awesome. She knows everything about vectors and I had the privilege of learning about vectors from her. There was just so much information; lentiviruses, retroviruses, promoters, suicide genes, backbones, and what all of the letters and numbers mean on her eppendorf tubes.

I finally learned what my project was the next day, 4XBDNF as I like to call it. What if I want to produce higher amounts of protein? What are the options? Well higher MOI’s (more virus per cell) could be used, but the FDA won’t allow that because it could cause problems like insertional mutagenesis and cancer. So other options? Why don’t I just add additional copies of a gene to a single virus? So that’s what I did. Because Brained Derived Neurotrophic Factor (BDNF) is a highly characterized protein in lab due to its importance in Huntington’s Disease research, this concept was based around the BDNF gene. I did all of my DNA work with the Goddess herself. I must of tried at least 7 times to get that stubborn cassette of 4 BDNF genes to clone into the vector backbone. It just wouldn’t go in. Finally it went in and I was so grateful because I finally got to start packaging my vector.

I began working with Catherine Nacey to package my vector, because a virus doesn’t work if its just a plasmid. I got culture cells, transfect cells, and eventually harvest my virus. This probably was the best part because I had created my very own vector.

I then needed to try it to see if it worked, right? So I tranduced some Mesenchymal Stem Cells with my virus to see how much BDNF it produced. AND drum roll please…… The cells transduced with my virus didn’t produce anymore BDNF than the wild-type cells (CRIES)!!!! Yep. It didn’t work and me, my mentor, and Dr. Fernando Fierro all have no idea why it failed. It kinda sucks it didn’t work, but what can you do.

I just want to say thanks to all of the awesome people in the Nolta Lab, Dr. Jan Nolta, Dr. Gerhard Bauer, Catherine Nacey, Kari Pollock, Dr. Fernando Fierro, and the Goddess for everything.

(P.S. I guess I’ll have to come back next summer to fix it!)

Alfonso Barraza

Alfonso submitted this video about his experience:

The New York Times and all the knees that are fit to print

Photo by Keith Kissel: 3D printer making a plastic turtle

Most of the time I’m just happy if I can get a document from my computer to print out on the nearby printer, so I’m always amazed to read about people using what are essentially modified versions of these printers to create 3D models of coffee cups, guitars, even ears. That’s why a recent New York Times article about a CIRM-funded researcher, Dr. Darryl D’Lima of the Scripps Research Institute in San Diego, who is using a 3D printer to try and create some spare knee cartilage caught my eye.

Dr. D’Lima got more than $3 million from the stem cell agency to research the potential use of stem cells in cartilage regeneration and treating osteoarthritis (you can read about that here)

The 3D printer project wasn’t part of that particular grant but the research the money funded clearly helped pave the way for the experiment.

The article does a great job of exploring how this kind of approach – using 3D printers to create all manner of materials – is becoming increasingly common in the life sciences, but it also points out that it is a long way from producing living tissue that acts and behaves the way that the real stuff does.

The reporter quotes Brian Derby, a researcher at the University of Manchester in the UK, saying:

“Nobody who has any credibility claims they can print organs, or believes in their heart of hearts that that will happen in the next 20 years.”

Dr. D’Lima admits there are a lot of obstacles to overcome but says when it comes to making cartilage he is cautiously optimistic.

“Printing a whole heart or a whole bladder is glamorous and exciting, but cartilage might be the low-hanging fruit to get 3-D printing into the clinic.”

If it works it would be a gift to the millions of people who currently undergo knee surgery every year to repair the damage caused by worn-out cartilage or from people suffering from osteoarthritis. But if we have learned anything over the years it is that stories like this make for great headlines, but often there are years and years of hard work before they make real therapies.

Kevin McCormack