Results
This page includes two sections: The story of my work over the last 2 years since I got the grant, including achievements, challenges, and pictures. The second section includes advice for future plant tissue culture amateurs, especially young people who want to or need to start at home on a budget. Thanks again to everyone who supported this project!
The story
As it's also noted in the results abstract, getting lab space as an undergraduate student, was unfortunately the major constrain for this project. Though I'd got a "yes" from Dr. Cecilia Franco, head of the Biotechnology Engineering department at my hometown campus, even before I entered college (and before I even applied to this grant), and I insisted multiple times on getting dates when I would be able to work at the lab, her responses were never clear.
Meanwhile, I got basic equipment and reagents to do some experiments at home, including an incubator, well plates, growth hormones, a working table, and MS media. I paid a PhD and a Masters student to train me on the basic tissue culture techniques, and how to potentially adapt these to a home lab to prevent contamination.
After making dozens of different contaminated flasks, and since the funding was not enough for a basic safety cabinet, I paused the project in November 2022 until I was able to get lab space at my university's larger campus in Monterrey, which I communicated with Breanna Duffy who was leading the New Harvest side of the grant.
In December of 2022, I contacted Professor César Puente from Campus Monterrey, who kindly agreed to mentor me so I could continue with my project in Monterrey. He took pat in redesigning the project, explaining that I needed to grow my own plants from certified seeds in the school's greenhouse before I cultivated anything in-vitro, and then culture shoots in-vitro before culturing anything else, to ensure that I could use the right experimental protocols, characterize my results correctly, and prevent more contamination issues.
We couldn't get permission to use the greenhouse until November 2023, but it was finally in April 2024 that I was able to successfully establish a in-vitro culture of cotton shoots from axillary buds of the greenhouse plants. By that point, I'd co-authored a paper analyzing the economic challenges and opportunities of developing cellular agriculture in developing nations... and (unrelatedly) my professor said he wouldn't support the initial project I proposed to him and for which I'd got this grant.
Advice for plant tissue culture amateurs
As of December 2024, it's clear that the initial proposal failed. I did not make a cotton trichome cell line, and I could barely even culture some shoots until years later. If I were to start over again, or advice someone (especially a young person) who wants to do plant tissue culture, plant cellular agriculture, or plant synthetic biology experiments at home, I would advice the following:
Sterility is the number one thing to achieve first.
a. Invest in a safety cabinet. You can get one for less than $500 US. I'd also recommend using an alcohol lamp inside it to carefully sterilize your tweezers.
b. Use Parafilm and, preferably, tissue culture vessels instead of Gerber flasks. You can get most other tissue culture things, including reagents, from PhytoTech labs.
c. Autoclaving everything you'll use during inside the safety cabinet (tweezers, flasks, petri plates). Use an InstantPot (or equivalent) and do not use it for anything else apart from your experiments. Use indicator tape to know you autoclaved for long enough.
d. Buy a working table (metal suggested) and work in a room where only your experiments will happen. Kitchen labs are by no means made for plant tissue culture.
e. It's easier, ecofriendlier, and safer (if you're using an alcohol lamp) not to use gloves of any kind. Sorry, you'll have to constantly rub your hands with a lot of alcohol every time before your hands go into the safety cabinet.
f. Watch videos from Plant Cell Technology and Plants In Jars on how to get started doing plant tissue culture at home. Both got great tips and ideas.
Especially when you're doing these types of experiments (or any kind of lab work at all) for the first time, planning is essential. Your research proposal is likely not specific enough for what you'll actually do in your experiments. You need to:
a. If you've never done experiments like this before, you want to do a "Hello world" of sorts. Making contamination free media, germinating an "easy" plant like a beans or lentils from their grains, or maybe growing an axillary buds are good places to start. This will force you to go from the theory you've read in the papers or conversations, to actually practicing the basic techniques, workflow, instruments, equipment, and even tricks you'll need for more advanced experiments.
b. Create a database for the different protocols you check out in the literature. Include the media composition, temperature, humidity, equipment used, light, pH, and other growth conditions each paper mentions. Equally important, include the results each study got, like germination rate, dry weight, or contamination rate. Your decision on what protocol to follow should be based on this data, and what you want to achieve.
c. Write down your protocol (materials, reagents, equipment, steps, considerations) on a physical notebook (yes, works better than typing and helps decrease contamination risk). You can use this same notebook as a lab journal to write down notes on things that went unexpectedly, tips someone like a mentor gives you to improve your technique, and (very important) mistakes you make, especially those that are stupid enough for you to forget over and over again. For example, I would write down things like agar concentration and whether a phytohormone was autoclavable.
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