“… I have grown motivated and determined to pursue a career in the sciences, but more importantly, to have my work make a difference in the world.”

By: Hector Medrano

In a mere blink of an eye, four weeks have passed since I first stepped foot in the laboratory, and dove straight into the experimentation that is being conducted under my wonderful mentor, Dr. Vaithi Arumugaswami. Over the course of the past four weeks, I have worked alongside each of my laboratory members, witnessing the unique approach they each put forth toward tackling healthcare’s greatest challenges. Reflecting upon my time in the laboratory, I can proudly assert that I was placed in one of the best laboratories at the Cedars-Sinai Medical Center, Board of Governors Regenerative Medicine Institute. Such certainty is derived from the numerous hours that my fellow laboratory members have taken out of their terribly busy schedules to instruct me on how to perform different procedures while explaining the science behind their experiments. What was most exciting was when I was given my own plate of 25i induced pluripotent stem cells (iPSCs), further allowing me to learn how to properly coat six- well plates with Matrigel, plate and clean the cells, change their media, and split the cells after becoming too confluent. Each day, as I changed their media, and observed their confluency under a microscope, I stood astounded as I viewed the gradual development of these cells. However, my experiences did not end there, as I was able to attend several conferences with renowned speakers, classes with professionals in their field, and tour world class facilities. For such memorable/breathtaking experiences, I must thank Cedars-Sinai Medical Center, as well as the California Institute of Regenerative Medicine, for providing a program where high school students are exposed to groundbreaking research. Observing the intricacies revolving around the world of research, and the passion and excitement with which my laboratory members come to work, I have grown motivated and determined to pursue a career in the sciences, but more importantly, to have my work make a difference in the world.

“This week was especially memorable – I feel like I learned so much!”

By: Soah Franklin

Week four of CIRM! I can’t believe we are already halfway through the program – it feels as if time is flying by. This week was especially memorable – I feel like I learned so much! On Monday I started my project, which I will present on Poster Day in a few weeks. I will be observing and documenting the effects of microRNA on corneal epithelial cells. I have been reading background information on many of the techniques that we will be using, like western blotting, immunostaining, and immunohistochemistry. The highlight of my week was observing tissue culture experiments, and learning about the process of coating the plates and seeding the corneal epithelial cells. Our lab is working with miR-146a, which has been shown to be involved in corneal epithelial homeostasis. This week we have been preparing my experiment: coating the culture plates with collagen and seeding one hundred thousand corneal epithelial cells per well in a 6-well plate. The cells will be transfected with miR-146a or the scramble miRNA as a control today. On Monday we will process the effect of miR-146a in the treated cells by immunostaining. I am so excited to see our results!

“…filled with learning the technicalities and maneuvers of different procedures, equipment, and computer software.”

By: Emma Friedenberg

This week in the Mattis Lab at Cedars-Sinai Board of Governors Regenerative Medicine Institute was filled with learning the technicalities and maneuvers of different procedures, equipment, and computer software. The most interesting and hands-on experience was RNA isolation, the purification of RNA from biological samples. The isolation of RNA with high quality is a crucial step required to perform various molecular biology experiments.

Ribonucleic acid is a polymeric substance present in living cells and many viruses, consisting of a long single-stranded chain of phosphate and ribose units with the nitrogen bases (adenine, guanine, cytosine, and uracil), which are bonded to the ribose sugar. RNA is used in all the steps of protein synthesis in all living cells and carries the genetic information for many viruses.

A rapid method of analysis for determining the identity of neural stem cells (NSCs) and their sublineages involves the early detection of differentiation markers tracked at the RNA level. This week, I learned the procedure of RNA isolation from cell lines. The first round of RNA extraction was performed on a small set of three cell pellets – one each for my two mentors and one for me. Each step was rotated between the three of us for learning practices. The next day, a total of 18 cell pellets of our cell line underwent RNA isolation which was executed by myself and my post-doctoral mentor. By the end of the procedure, 1 microliter of RNA was transferred to the computer analysis system. Fortunately, all of the cell lines achieved an RNA concentration above the contamination or error mark.

“….there is more to researching than pipetting and extracting RNA-I say this because it has been big this week but rather it is a lifestyle for many individuals.”

By: Dennis Portillo

As the third week of the CIRM/SPARK program nears its end, the days that have passed have almost been a blur. The routine work of being in a lab has now been established, but still each day a little bit different than the previous. That statement sums up the lives of most scientists, who do not follow the standard procedure of simply working five days a week and taking holidays off. The life of a researcher, in particular, is not one that is necessarily easy, rather but be a journey of ups and downs, sprinkled in with blood, sweat, and tears (just not in the experiment hopefully). From this week, the task of being a researcher goes far and beyond more than just the occupation in its name, with experiments at times taking longer hours than most are accustomed to, and any mishap having the possibility of setting it back experiments. Perhaps learning this lesson in itself is more memorable than any sort of hands-on bench work, or anything that could be attained from an experiment. Talking with my mentors this week has helped me realize that there is more to the questions of simply why and how, using these to stem into other more valuable questions as well as accepting the fact that experiments are not as straightforward as one may think. While it may take a few minutes to a few hours for all the procedures to be done if lucky, it takes much longer to even analyze what has been done and gained from it.

While the past two weeks have mostly consisted of collecting and analyzing data, it has something that I’ve come to enjoy. For many, the summertime has become one of relaxing and enjoying, but the same is not necessarily true for scientists. Earlier in the week, I was discussing how many are eager to go into the field, yet not prepared for much of what they have to give up, with sometimes the future exchanged little in success. While saying this is by no means an effort to be cynical, it gives hindsight to have a realistic point of view and how the main motivation as to why you get up in the morning and power through the day being passion and drive. Perhaps a few cups of coffee may also do the trick but that’s not the point. For many scientists, their work is their everything, and even if the original hypothesis may turn out wrong, or the data provide very little, many falls in love with their work (and rightfully so). So, while counting cells and looking at neurons under the microscope may be some notable highlights of the week, the real takeaways that come from this week are the ones that stay with you forever. As cliché as that may sound, there is more to researching than pipetting and extracting RNA-I say this because it has been big this week but rather it is a lifestyle for many individuals.

“…the knowledge I have acquired and the friendships I have already developed are unlike anything I’ve ever experienced, and I cannot wait to continue my work in the weeks to come!”

By: Candler Cusato 

I can’t believe I’ve already been in the lab for two weeks; it sounds cliché, but time certainly does fly when you’re spending almost seven hours a day learning, experimenting, and meeting new friends. Overall, this week has been filled with simultaneous challenges and triumphs, stretching me as both a scientist and an individual, which I can’t wait to share.

After I toured more of the Cedars campus on Monday and Tuesday, I attempted the most challenging scientific technique I’ve ever been exposed to: retinal whole mounting. Over what seemed like days (but was probably only an hour and a half), I removed the cornea and lens from a rat eye under a microscope and detached the retina from the rest of the eye casing to mount it on a slide. This technique allows you to view the entire retina while preserving vascular and retinal structure in your specimen, which cannot be accomplished by other sectioning techniques that involve cutting tissue with a machine. To understand the difficulty of removing a PBS-soaked retina from the remains of an eye, I would compare it to trying to turn the pages of a soaked, 1000-year-old book without ripping the pages. In short, it is VERY difficult, and it will definitely take me much more tries to improve my technique.

Outside of the lab, I was thoroughly enthralled during my lunchtime learning sessions this week, which is humorously deemed “Knowledge Noshes,” particularly during the presentation by Dr. Robert Baloh, MD, Ph.D. His progress in creating gene and stem cell therapies for amyotrophic lateral sclerosis (ALS/Lou Gehrig’s Disease) is truly fascinating, and his passion for the subject was palpable during his lighthearted but educational presentation. He peaked my curiosity in neurology so much that I am intrigued to explore more about the subject of neurodegenerative diseases and possibly pursue it in my future studies.

Also, this week, I gained a new laboratory companion, as Amanda, an intern from Brown University, arrived on Tuesday to join Dr. Wang’s lab. In just a few days, we’ve braved whole mounts together and suffered through the slow-loading microscope camera software while waiting to take photographs of retinas. With Dr. Wang’s help, we are on our way to perfecting our labeled retinal images in Photoshop and learning the various eye histology techniques and the intricate system of blood flow throughout the eye.

As this second week of CIRM Spark comes to a close, I feel even more fortunate that I was selected for this prestigious opportunity, as the knowledge I have acquired and the friendships I have already developed are unlike anything I’ve ever experienced, and I cannot wait to continue my work in the weeks to come!

“I am slowly yet surely unraveling and participating in, and I look forward to all that is to come.”

By: Kevin Kim

It’s week two already, and I cannot believe how quickly the lab experience has turned up its pace! I’ve finally participated in some hands-on laboratory procedures while reading lots of journals and papers on corneal epithelial cells and wound healing – the life of a researcher has many facets, it seems.
One of the highlights of this week was thoroughly learning about and performing the ICC, or immunocytochemistry, staining, which localizes antigen expression and is dependent on specific epitope-antibody interactions. ICC uses cell cultures and generates a strong and specific signal for each antigen of interest. For example, detection of an abundant protein in cultured cells may require a short fixation period, minimal blocking, and may be compatible with direct visualization using a fluorochrome-conjugated primary antibody. In contrast, detection of a phosphorylation-dependent epitope in a section of archival tissue may require antigen retrieval and be dependent on amplified chromogenic visualization.
The procedure is a five-hour process, so I spent morning to afternoon working on this part of the lab. Although a seemingly daunting and arduous task, the ICC was quite simple with the help of the Research Associate, who carefully and expertly explained each step while having me simulate and perform the exact actions and procedures.
I never really understood why there are so many individual lab procedures involved in the grand scheme of things, and I didn’t quite grasp how all of these facets related to the lab’s focus on gene therapy and corneal diabetes. However, that is a question that I am slowly yet surely unraveling and participating in, and I look forward to all that is to come.

Candler Cusato

Blog 1 Entry:

Hi, my name is Candler Cusato, and I am rising senior at Los Alamitos High School in Orange County. But, more importantly, I am one of eight high school research interns selected to participate in seven weeks of stem cell research funded by the California Institute for Regenerative Medicine at Cedars-Sinai Medical Center. For the next two months, I will be exploring the progression of retinal degeneration and stem cell-based therapies to combat blindness in the millions of patients suffering from retinitis pigmentosa and age related macular dystrophy, under the direction of Dr. Shaomei Wang of the Regenerative Medicine Institute, and this is the account of my first few days as an intern in a state-of-the-art scientific lab!

After waking at 5:45 AM and braving the daily 405 shuffle to get to Cedars from Long Beach, I arrived on Monday morning and immediately fell back into my usual pattern of rapid note taking during biology lectures, as program director Dr. Virginia Mattis gave us an introductory lecture on stem cells. With her help, my eyes only slightly widened when I reached the lab later that afternoon and everyone uttered acronyms, such as IPSC’s, IVF, FDSC’s, with practically every breath. Luckily, they fed us pizza to keep our heads from spinning, and I went home feeling proud but slightly cross-eyed, after I spent my afternoon gazing into the microscope and successfully removing rat lenses and corneas.

Tuesday morning, after the slight hiccup of waiting for the wrong elevator for approximately 10 minutes, I was immediately whisked into a meeting between Dr. Wang and her outgoing intern, and I listened attentively to their conversation on how human cell injections influenced rat RPE production in the retina. Searching, as all scientists do, for more data, we all journeyed into the lab and examined the retinal slides tagged with various fluorescents to show the distinction between cell types, seeing first hand how the injection of foreign cells significantly decreased retinal pigment epithelium. By the end of the day, I had also helped feed and change the medium in the lab’s neurosphere cultures, which I learned will soon become my full responsibility, and journeyed down to Third Street to discover the wonders of Somma Somma, a Chipotle-style Mediterranean restaurant.

Wednesday morning, after a long conversation with my mentor about my life and college plans, I went to work preparing slides and examining them under the microscope to search for folds in our eye section samples, easily able to be ruined since they are only millimeters thick. With help from the post-baccalaureate intern, Alice, I was able to identify the layers of the retina, as she walked me through their usual structures and appearances under the microscope after being stained with cresyl violet. Thursday, I continued my work in staining, learning the complex and ten-step process that ensures my hard work does not go to waste and my slide contents can be seen under the microscope. Although my first batch was too light, I eventually was able to view my specimens, and to my immense relief, they were identifiable as retinal cells. Score!

In days at Cedars so far, I have been surrounded by unparalleled friendliness and pursuit of knowledge for the overall benefit of mankind. My time here, although brief, has already yielded a new understanding of eye structure, disorders, and laboratory techniques crucial to my success in my future undergraduate studies. If this week is any indication, I am confident that I will leave in August with a newfound appreciation and understanding of scientific research and the preparedness to undertake a career in the scientific field.