Stem Cell Research
Investigating the effects of naturally occurring electrolytes and commonly prescribed heart medications on human induced stem cell derived cardiomyocytes
Aspirating medium from culture plate CARLOW Center for Medical Innovation, Pasadena 2023 (Trial 2)
Poster Presenter at Archer STEM Symposium Archer School, Los Angeles 2022
Serological pipetting solution during immunocytochemical cell staining (Trial 1) Pathways to Stem Cell Science, Los Angeles 2022
Examining the antibody staining results on the Fluorescent Microscope (Trial 3) CARLOW Center, Pasadena 2023
Poster presentation at the International Society for Stem Cell Research Symposium Sao Paulo, Brazil 2023
Virtual presentation at the Society for In Vitro Biology Research Meeting Norfolk, Virgina 2023
Heart disease is the leading cause of death in the United States, and cardiomyocytes are one step closer to less invasive research methods and treatments for cardiac ailments. The goal of trial 1 of experimentation was to determine if the heart supplements added before cardiomyocyte development would impact the differentiation process and cell yield. Differentiating human induced pluripotent stem cells (hIPSCs) were treated with the supplements L-Carnitine, Coenzyme Q10, and Omega 3 Fatty Acid. Differentiation efficiency was then assayed with NKX2.5, a cardiac differentiation marker. Omega 3 fatty acid had the most fluorescent color, thus had the most amount of cell differentiation. This trial answered my question of which supplement causes differentiation.
The human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) can be used to model cardiac pathophysiology and personalize medical treatment on a case-by-case basis to yield better patient outcomes. However, the biggest challenge is that the hiPSC-CMs remain in a stage resembling fetal cardiomyocytes rather than adult. Therefore, for trial 2 of experimentation I decided to investigate which medications and electrolytes impact hiPSC-CMs maturation and cell growth. TBX5 is a key transcription factor necessary for the development of cardiomyocytes and the formation of the cardiac conduction system as the cells mature. By measuring the expression of TBX5 during the induction of HiPSCs into cardiomyocytes, we can track the effects of various treatments on the ability to allow for the formation of mature cardiomyocytes from HiPSCs. The results of this trial were that each of the concentrations of electrolytes and medications impacted cell maturation and yield as seen by the antibody marker and results from the phase contrast microscopy. 10 mM (millimolar) of metoprolol and 10mM of Potassium chloride show to be the most promising supplements for health and robust hiPSC-CMs.
The last trial of experimentation combined aspects of trial 1 and trial 2, but still investigated cardiomyocyte maturation. This trial used the electrolytes potassium chloride, magnesium chloride, calcium chloride, and sodium chloride. It also used the commonly prescribed heart medications metoprolol, digoxin, hydrochlorothiazide, alpha lipoic acid (omega 3 fatty acid), and the health supplement L-carnitine. This trial also tested each of these additives at differing concentrations 25mM, 50mM, and 100mM. The results from the supplements from the previous two trials were confirmed in trial 3. Overall, Digoxin at 25mM and 50mM concentrations had the greatest amount of mature cardiomyocyte staining. Calcium chloride at 25mM concentration followed in the greatest amount of maturation. The cells that were treated with CaCl had the largest cell yield 25mM, a lower yield at 50mM, and the lowest yield of this batch at 100mM. This shows that higher concentrations of calcium inhibits cell growth and maturation. L-carnitine at 25mM, 50mM, and 100mM had slight antibody signals indicating that it had a limited amount of mature cardiomyocytes. The cell wells of L-carnitine at Day 15 also had a lower and more spread out yield under the microscope. These results can be explained by the fact that L-carnitine has several possible destructive properties. L-carnitine is a metabolic enhancer that oxidizes fatty acids to create energy. The excessive burning of amino acids, as well as the production of too much energy, would explain why L-Carnitine was not successful, especially at higher concentrations. Overall, this trial was very positive with Metoprolol, calcium chloride, and sodium chloride showing the most promising results.
For the upcoming 2024 Los Angeles County Science Fair I will be submitting a project that explores how the Covid-19 spike protein impacts the electrical impulses of cardiomyocytes derived from induced stem cells.
Research Summary
A newspaper section in the Pasadena Star-News
Additional links
Highlighted on pages #4 & #27
Covid-19 has major impacts on the physiology of the human body, namely the body's cardiopulmonary systems. Investigating the effects of this disease on the heart at its most basic cellular level will help scientists further understand and analyze potential underlying concerns with this virus. If the covid 19 spike protein changes the structures and potentially the functions of these cardiomyocytes then it can explain some detrimental effects of the disease or help scientists discover the nature of the sickness further. Fully grown cardiomyocytes that have been differentiated from human induced pluripotent stem cells will be transfected with recombinant Covid-19 spike protein. The transfection procedure consists of proteins being directly put into cell bodies using a lipid reagent. The cells will then merge within a few hours and change accordingly to the spike protein (or remain unchanged). The results will be measured using a immunocytochemical procedure that observes changes in cell morphology using an emerald fluorescence protein. This antibody staining will allow for the visualization of changes in cell shape and size.
Los Angeles County Science and Engineering Fair 2024: Current Research
Girl Scouts Gold Award
The two root causes I addressed with my gold award work are (1) The attitude of women in STEM and trying to break the concept of women not being successful in STEM fields and (2) stigmas towards mental health and how to address it and cope with mental health issues.
I did a series of STEM-related workshops at the Crittenton facilities as planned. I worked with three groups of children with a range of 16-21 children in each group. I started with a presentation on the scientific method, discussing the different types of sciences we learn, and talking about famous women pioneers in science. I followed these discussions with demonstrations of the different experiments we would be completing. All the children had the opportunity to perform each experiment, which was a wonderful learning experience for them all. This hands-on experience definitely added to their science education and possibly sparked an interest in the possibility of future careers in science despite not having much exposure to the subject beforehand. I think all the groups most enjoyed the density column as both the hands-on learning and visual aspects excited the girls.
As planned, I addressed mental health issues by doing a dance and mental health workshop for the five cottages at the Crittenton facility. Each girl received a package of items that could help in stress relief: stress balls, face masks, pencils, and notebooks to write their own feelings and hand lotion. I also described how movement, specifically dancing could help in managing stress. I am an Indian classical dancer of the Odissi dance form so I danced for each group and taught them a few steps as we danced together. The girls were delighted and stress-free as we performed in groups.
Those who are housed in this organization have been separated from their families due to multiple reasons. Some have left their country to escape war or to find security and opportunity in America. I tried to find ways to help them during this stressful time in their life. I focused on their mental health and made them feel at ease and secure. Most of the time children in such stressful situations struggle with prioritizing their mental health and education. This is a global issue I would like to address. As a teenager living in a secure family with access to all the resources I need for my education I always felt the urge to help those who are deprived of it. I also addressed the issue of the dearth of STEM education and representation for women/girls, which is a global problem.
Performing for the girls
Teaching group 1 dance steps
Teaching group 4 dance steps
A picture with my project advisor, Ms. Berryman
Girls observing the strawberry DNA extraction solution
Speaking to the presentation on female scientists and the scientific method.
Divya Jyothi
Photos from volunteering at the Divya Jyothi Charitable Trust for the Disabled in Mysore, India https://divyajyothitrust.org/
Presenting the tactile globe I donated to the trust
Talking to the girls
Observing the process of creating the braille books
Printing the book
Girls learning in their geography class
Conversing with the principal of the institution
In this activity I was asked to sing a song in English to the girls in their classroom. I chose to share the song "This Land is your land" due to its strong and pertinent message. I really enjoyed teaching then singing with the girls, and seeing the smiles on their faces as we all sang together.
Odissi Classical Dance
As an Odissi Classical Dance performer I have practiced and perfected the technicalities of this art form for over 12 years. Please click the video below to view my showcase of this dance form
Certifications
In order to further my stem cell training, I participated in these training courses hosted my STEMCELL Technologies. I learned valuable techniques specifically for stem cell dervided neural cells to prepare for future neural induction experimentation.
I really enjoyed my AP Calculus AB course so I decided to explore real-world applications of the subject.
After several History Day and Science Fair competitions, I wanted to retrace the steps of medicine and delve into the beginnings of medical practices.
In the summer of 2021, I took a Research Methods in Psychology class at the Pasadena Community College. The course required knowledge in research statistics. So prior to the beginning of the class I completed this statistical thinking class in preparation for the college psychology class.
Occidental Research
As a research assistant, I have learned important skills for field research. This journey has not only provided me with a deep understanding of the processes that shape a child's mind, but has also allowed me to investigate how environmental factors impacts a child's growth. By conducting in-depth interviews and observations I've uncovered valuable insights into of the cognitive and intellectual milestones that mark child development.
Humanities Competitions
California State History Day Competition in Sacramento, California 2022
Los Angeles County History Day Individual Exhibit Category Finalist 2022
Los Angeles County History Day Individual Exhibit Poster 2023
Scholastic Art and Writing Contest Poem 2022 - Result: Silver Key
The endless expanse of the deep ocean filled almost
Every inch of the picture he painted in his head, as that was all he saw for weeks on end
The faces of those he loved loomed as the storm clouds did above
The idea of ever returning to them kept him alive
But for how long?
How long must he stay
A painting holds thousands of words they say
The dark ocean that surrounded the old wooden boat only remind him of the screaming
It only reminded him of the pleading, crying, and hysteria
It only reminded him of the people he could not save, of the friends he could not save
Why does the cruel world have the authority to take one's life?
Why do our oceans and mountains cause the downfall of human life?
Why can’t he live his life peacefully?
Why must he stay in this wooden boat in the middle of the vast ocean day by day?
A painting holds thousands of words they say
Why was he chosen to be the last to survive?
To suffer
To bear with the pain of guilt
The pain of loneliness, shame, and dismay
A painting holds thousands of words they say
The crashing waves and the screeching of the vultures rung in his ears
How come the vultures had a home
Had a destination and family to go back to
Even they found their way
A painting holds thousands of words they say
What did he have left?
If going to the promise land meant losing everything he had
What was the point?
Could the promised land keep its promise?
Or will he continue to suffer in pain
If going to the promise land meant meeting eye to eye with death
Meeting eye to eye with misery
Then he did not want it
This is what he thought as he was led astray
A painting holds thousands of words they say
The picture in his head would only be ocean
The distant memory of himself was lost within its depths
This is what he chose to display
A painting holds thousands of words they say
So the man with the painting lay
But the truth is this painting only holds one
And therefore his time was done
Thousands of Words
A dash of cardamom
A sprinkle of salt
A handful of spices
The most potent is love
Every dish makes a statement
A statement of pride
The silks and the cotton
The beads and strings
Hand sewn together
In intricate patterns
Each sari flowing in the wind
The achol waving as if saying hello
The music filling the air
The different dances
Every song significant
Having a purpose
The music plays
The people take to the streets
Laughing and smiling
As their anklets jingle
Each foot placed on the right beat
Twirling and moving about
The overwhelming joy
As they dance the night away
No worries about tomorrow
The festive traditions
The colors
The lights
The brother and sisterhood
A connection to the past
A link to the history
These are the meanings of India
Scholastic Art and Writing Contest Poem 2021 - Result: Honorable Mention
Land of Beauty