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RMI CIRM Spark

“The greatest aspect of this internship is .. everyone genuinely loves what they do”

Submitted by Jennifer Siddique

I fear becoming too accustomed to the daily life of working in the Svendsen Stem Cell Research Laboratory only for it to come to an abrupt end. For this reason, I am making the absolute most of these last few weeks.

Everything I do in the lab is a first for me. I am very fortunate that my mentor, Dr. Veronica Garcia, is an electrophysiologist as I get to to become acquainted with electrophysiology. Prior to this internship, I had absolutely no idea what electrophysiology was. I recall the first time I met my mentor and she questioned if I had any knowledge of electrophysiology. My response was that I have never even heard of the field before. [Sorry, Veronica.]

Earlier this week, I obtained a brief glimpse into the world of electrophysiology. Dissociated neuronal cultures are grown on microelectrode arrays (MEAs). Microelectrode arrays are devices used to measure unpremeditated activity of neuronal cultures. I learned how to operate an MEA. A few of my findings can be seen on my poster at the end of this internship.

Perhaps and is kind to one another. I have always struggled with social anxiety. That is, I am constantly overthinking and overanalyzing social interactions. It is quite the battle. Fortunately, I have built a lot more social confidence as a result of everyone’s welcoming personalities and overall desire to help others. I have also made a lot of new friends in the lab.

 

With that said, I am still in disbelief that I somehow attained this stem cell research opportunity. I am forever grateful for this experience. Not a day goes by where I have not learned something new in the lab.

A Perpetual Growth in Knowledge

Submitted by Andrew Choi

The fact that I was able to learn so many new concepts throughout the close-to-five weeks I have interned here at Cedars Sinai seems to be very surreal, mostly thanks to Gen! There were new and, as I would regard them, very valuable occurrences since my last blog update as an intern in the CIRM Research Program.

During the first two weeks, I read and annotated scientific articles so that I can assimilate, although not entirely, with many of the techniques that my mentor has used throughout her career. These articles and scientific journals that I read did not solely pertain to techniques such as stereology, but also background information, which I found to be quite helpful throughout the latter weeks of my internship.

I’d say that my studies for the coming weeks are continuations of preliminary findings and data gathered by my mentor. My mentor’s research project is majorly based on amyotrophic lateral sclerosis (ALS), a devastating disorder causing detriments to lives across the globe. ALS is a disorder or a condition in which motor neurons controlling muscle function degenerate leading to paralysis and ultimately, death.

My research project that I will be focusing on this summer is the effect of neural progenitors engineered to produce growth factors on degenerated motor neurons. In order to observe the experimental data, my mentor and I performed a technique called immunocytochemistry (ICC) where we treated neuronal stem cells with certain antibodies such as Stem121 (recognizing anti-human cytoplasm) and recognizing various growth factors. Thinking back to the time when I performed ICC with my mentor, I am very grateful for my mentor’s patience (haha!)!! I broke several coverslips with the forceps here and there while trying to prepare the slides. We incorporated the MBF Stereology software to our protocol to quantify our qualitative data. These techniques enabled me to count and characterize cells expressing certain markers so that I can evaluate the differences (if any) in the differentiated cells, neurons and astrocytes. I don’t necessarily consider myself to be decent at these techniques, but rather a pretty experienced amateur.

You may think that staring at a screen for several hours and counting cells with stereology, may have drained my eyes and enthusiasm, but the excitement I have for the program continues to thrive! I become more excited every day to know that my mentor has new concepts and life lessons to teach me!

“I am meeting so many great People .. they are teaching me so much”

Submitted by Jeremiah Fountain

Over the past 3 weeks, I have loved working in a lab at the BOG Regenerative Medicine Institute at Cedars-Sinai Medical Center. I am meeting so many great people and they are teaching me so much. I have sat in about 6 meetings and in there I think I learn the most about my research and others projects in the lab.

My first couple of weeks involved several hours of orientation and safety information. When I finally got into the lab my mentor Anjoscha Kaus taught me the basics about her project and what she does. She is focusing on Amyotrophic lateral sclerosis (ALS), a progressive neurodegenerative disease that affects nerve cells in the brain and the spinal cord. ALS damages the motor neurons (MNs) throughout the body. Upper MNs connect from the brain to the spinal cord and lower MNs from the spinal cord to the muscles through the body. The ability of the brain to initiate and control muscle movement is lost when the motor neurons die. Some people may lose the ability to speak, eat, move and breathe because of the loss of voluntary muscle movement that was affected.

The researchers here identified metabolic signatures in MNs they generated from C9orf72 ALS patient-derived induced pluripotent stem cells (iPSCs). The C9orf72 gene so far is the most frequently found gene connected to familial ALS. In their metabolic analysis the team here found a number of protein levels to be different, among those Ornithine Aminotransferase (OAT), an enzyme necessary for Proline synthesis, and Glycine Aminotransferase (GATM), critical for creatine synthesis. Both have been found to be dramatically increased in ALS samples, as showed by means of Protein detection through Western blotting evaluation. The 2 proteins closely intersect as GATM produces ornithine as a byproduct that is the essential substrate for OAT to synthesize Proline. This difference was accompanied by increased amounts of tRNA ligases, which are required for synthesis of RNA and proteins. Now we investigate in what CNS cell types the two candidates are expressed in ALS iPSC-derived motor neurons.

I am very eager to finish off the last couple of weeks as a research intern and travel to Berkley with my peers.

 

 

Huntingtons Disease

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!

“I made my mistakes. I had my moments of glory”

Submitted by Nikhil Chakravarty

So, a lot has happened since I last wrote. Well, to start, I did a lot of lab work. Immunofluorescent Assays, Western blots, Gel Electrophoresis and all kinds of cool stuff. Doing all of this work has really made me appreciate the work and time that it takes to get publishable work done. All the concepts present in our lab education are some of the newest, most innovative concepts in medicine. Although having this opportunity to really further my knowledge is a dream come true, I cannot really say that this is my most impactful experience here.

I think that the greatest thing that I have done is create new relationships within the lab. Not just with my fellow interns, but the students in my lab and my mentor as well. I have formed friendships that have surpassed the simple ‘hello’ or ‘what’s up’ in the lab. We talk about all kinds of things from which basketball player signed with which team, what religious holidays are coming up, who went with who to prom. All kinds of stuff. Now, this all is really uncomfortable for me to write, so lets move on to less emotional stuff, eh?

My mentor has done a lot with me to further my knowledge of these cutting-edge concepts. He has trusted me to edit some of his manuscripts pre-publishing and I actually gave a presentation to discuss my project at our weekly lab meeting. In the lead-up to this, the task seemed extremely daunting. But, as I did it, it all seemed to fall in place and my lab was extremely receptive to what I had to say. They helped me to improve my content and the overall presentation, not only so that I can concisely and effectively explain my project at the conference in August, but also to simply give an effective presentation in the sciences. This was a huge lesson that I loved learning so far.

Now that all of the relationships stuff has been addressed, let’s talk about the science-y stuff! I got stem cells recently and it has been so cool, but also really intimidating. I was given the task of maintaining these cells and cleaning away the differentiated cells. This was the daunting task. I had to try and clean away these cells while not taking away any of the stem cells! Now, for those of you who haven’t seen these colonies, the cells are in close proximity to one another. This makes this even harder than it may seem. Anyway, I made my mistakes. I had my moments of glory. I even made my cells fluoresce so that I could image them through a fluorescent microscope! That was super cool. Overall, this whole experience has been a hodgepodge of different tasks that seemed disjointed at first, but, as I really dug down into the nitty-gritty of my work, I learned that, in any science, not everything is as it seems initially.

“I can’t believe I only have 5 weeks left!”

Submitted by Eshanika Chaudhary – June 30, 2016

Alright, so I’ve got almost 2 weeks down!  Time is flying, and I hate myself for using that expression, but it’s just true– I can’t believe I only have 5 weeks left!  I love that I get to explore a lot of different areas (such as imaging, surgery, and lab work) and see how the whole Cedars bench-to-bedside idea is actually implemented.  I was supposed to also observe a procedure today related to my project, but was not allowed to enter the restricted area for imaging.  But on the bright side, speaking of my project, I got to discuss my project outline with my mentor on Monday!

In the lab right now I’m still mostly shadowing but I’m working hands on too which is nice to get used to everything, and I’m planning to start doing my actual protocols very soon.  We also had our first knowledge nosh event yesterday where we got to hear a lot about scientific posters from two very well known professionals!  We got to see examples and we also heard tips about how to present and such.  I’m a very design oriented person in general and it’s been my “thing” since I was 13 (or 12? I don’t really remember) years old, so hearing about all the design possibilities for posters got me even more excited about beginning mine.  I also love public speaking (I know, it’s weird), so thinking about presenting the poster as a performance of sorts also got me really ready to start on it.  I can’t wait to present at Berkeley!  And in terms of content, I know that I’m going to have so much to say.

I’ve already learned so much, and my friends are beginning to get tired of me going on and on about how innovative this work is!  If you want more information about the work we’re doing in my lab, you can check out my first blog or the Gazits’ laboratory website here.  Keep checking the blog to stay updated on everyone’s progress throughout the summer!  And also, everyone should definitely check out my CIRM instagram I made for this program @cirmspark_eshanika for some pictures of me doing science, especially because I’m going to be posting a lot more on there than on this blog (I won’t be doing another blog post for about a month).

Lots of love and I hope everyone is having as great of a summer as I am!

“… a second home for me”

Submitted by Michal Fernandez – June 30, 2016

It’s been a whole week since I’ve started my internship program. The days have been filled with the hums of centrifuges and the sounds of liquid nitrogen tanks, yet strangely, I enjoy these sounds- it’s almost like becoming a second home for me. Throughout the week I’ve been busy  reading several articles and scientific journals assigned to me by my mentor- all in regards to retinal degeneration and general/advanced anatomy of the eye.; surprisingly, it is far more complex than I have expected, but, that does not deter me.

For the past few weeks, I’ve mostly been involved with activities related to histology. I’ve observed and helped a histologist with his given assignments such as immuno staining and/serial sectioning, to a certain degree. In regards to the eye, specifically the retina, I’ve been studying the general aspects of degeneration such as AMD (Age-related Macular Degeneration) and the general concepts of how such diseases perform and atrophy the retinal layers, mainly the photoreceptors.

Dr. Shaomei referenced me a previous study of her interest in regards to treatment of AMD. The treatment was basically to inject pluripotent stem cells (stem cells that can change into any somatic cell) in order to replace the retinal pigment epithelial (RPE) cells to stop/delay the death of photoreceptors. I did learn about the cons of using stem cells however, such as ethical concerns regarding a highly effective line of stem cells, embryonic stem cells. These stem cells come from human embryos under the blastocyst stage so, it raises the ethical questions of harvesting a human embryo for stem cells. Currently though, I’m not really that involved with cell culture just yet, I’m still within the histology department of the lab, observing and helping to collect tissues and staining samples of interest.

I do enjoy it a lot in here although, having no access to the floor I’m assigned to daily, gets rather inconvenient at times. Nonetheless, this has been a rather fun experience for me, I’ve learned many different techniques and information regarding immunostaining and staining in general as well as getting an introduction to cell culturing and a deeper understanding about retinal anatomy.

Michael F & Dr. Wang
CIRM Spark Intern Michael Fernandez & his Mentor Dr. Shaomei Wang

Nikhil Chakravarty

Hello from my first blog post! My name is Nikhil Chakravarty. I am an incoming senior at Granada Hills Charter High School. My mentor is Dr. Vaithi Arumugaswami. Starting this internship through the CIRM Spark Program has been amazing. However, although this feels like a dream comes true now, I do have to admit that I was quite nervous both coming into orientation and when walking into the laboratory. In both instances, I felt that I was about to encounter people who would not be afraid to leave me behind if I did not understand something. But, I must say – I was pleasantly surprised. Not only did I discover that my seven colleagues were thinking just like me, but I was welcomed into the lab with open arms and was given several opportunities to expand my knowledge on the processes going on around me rather than simply watching it happen! The two students in my lab have already given me things to do to help them in their research and to prepare me for my own research. I have already watched the making of a gel for and the process of gel electrophoresis, a western blot, and an immunofluorescent assay in order to stain the cells.

So far, I have not really done much to pursue my own research, but my mentor and I have sat down to discuss the overarching theme of that work. We will be dealing with stem cell differentiation in the liver, specifically in the development of the bile duct with a genetic pressure. This genetic pressure leads to the fetus not developing or having an underdeveloped set of bile ducts. The goal of this project is to see whether it is possible to construct a bile duct from these cells. Obviously, this is still in vague terms, as I am still learning about stem cell differentiation and this specific genetic pressure. I feel that my primary goal in this program is to gain a strong understanding in the process of stem cell differentiation, different possibilities of stem cell therapies, and how to conduct proper research in a laboratory setting. I think that the opportunities presented through this program are really going to set all eight of us down a path towards success in the medical research field and that my mentor is going to give me invaluable lab experience that I doubt I could get nearly anywhere else. Having the ability to perform jobs pertinent to the experiments in the lab from the first day, as opposed to simply washing flasks or beakers for the students in the lab, has probably been the most pleasant surprise for me. While I did know that I would be performing research, I was shocked that they gave me actual bench work starting the first day.

This whole experience has exceeded all of my expectations and it has only been one day! I cannot wait to see what invaluable experiences I will gain throughout the rest of this program.

Jeremiah Fountain

My name is Jeremiah Fountain and I am a research intern working in a lab at the Advancement Health Science Pavilion at Cedars-Sinai Medical Center. When I first heard about this opportunity by my mom I was ecstatic to finally working in a lab. Also I have been looking for a chance to have a job and obtain work experience. Cedars CIRM SPARK minors in research program was the perfect opportunity for me to experience working in a lab.

My first couple of days involved hours of orientation, where we had a campus art tour, I meet my PI mentor Anjoscha Kaus, and got a brief introduction of stem cells. Stem cells are basically a simple cell in the body that is able to develop into any one of various kinds of cells (such as blood cells, skin cells, etc.) Everyone at Cedars is so kind and helpful with anything I need. It was such a smooth transition coming into the research program and for my first time working in a lab.

The employees at Cedars really take safety in the lab serious and we had hours of orientation dealing with safety protocol. Andrea Ravard is a very skilled safety expert that taught all the interns about the safety codes and all the hazards in the lab.

Throughout the second half of the week I have been working with Gen while my mentor was gone, along with my other intern Andrew. Anjoscha researches ALS and how patient stem cells can unveil diseases. But for now I am working with Gen and Andrew and we are looking at progenitor cells secreting growth factors for the treatment of ALS.

I am very excited for what lays ahead throughout the program.

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