With all the new cannabis research currently being conducted, it may be difficult for a grower to find a paper on cultivation that ticks all the boxes. New lighting methods, cultivation techniques, ways to increase metabolite content, and bigger yields are all areas where research can provide valuable insights. At the same time, some cannabis businesses must necessarily start with a literature review to develop SOPs and similar guidelines. This is particularly true for tissue culture operations.
Reza Farzinebrahimi, a biotech researcher working for a Swedish biotech company, has been exploring the setup of cannabis tissue culture trials. However, he encountered numerous challenges, from finding a legal source of seeds to navigating the regulatory hurdles of cannabis research in Sweden.
Finding the right tissue culture way
While some of these issues have been somewhat resolved, another major challenge has hindered him from conducting proper scientific research: the unreliability of cannabis scientific papers. "When it comes to papers on cannabis tissue culture, there are tons of papers with many different methods," he explains. "For instance, when you read about a protocol for rooting and shoot formation, you'll find papers suggesting different hormones and techniques. However, if you try to replicate the same protocol in your lab, it doesn't work."
One might argue that this is because the protocols are highly variety-specific. However, Reza notes that he sourced the same varieties referenced in some of the papers he reviewed, applied the same protocols, and still didn't achieve the results claimed. "I tried more than ten different papers and couldn't replicate the same protocols with the same results. There's a lot of misleading information out there, and it's a huge problem if you're trying to get into cannabis tissue culture."
Reza explains that, unlike other crops, cannabis tissue culture is a relatively new field, making a thorough literature review essential for anyone starting a business. This is just one of the significant hurdles in tissue culture. "The other issue is funding," he points out. "A cannabis tissue culture lab is very costly to set up, and securing funding is a big challenge. If you're an R&D company, you often need to rely on private sector sponsors to research what they ask for. Scientists then produce papers based on that research. At the same time, there are many conflicting interests when it comes to publishing the actual results, as cannabis companies may fear losing their competitive edge. That's why I believe there's a lot of unreliable literature on cannabis tissue culture that needs to be taken with a grain of salt."
Trial & error
That doesn't mean, however, that all literature on cannabis tissue culture is outright unreliable. Reza points out that good papers exist, but they are hard to find. "This means that if you want to set up a cannabis tissue culture business, you need enough funding to go through a trial-and-error stage. You'll need to develop your protocols based on a literature review that may omit certain details or even results."
Reza also highlights a couple of red flags that may indicate a research paper is misleading. "Take the cannabis seed, for instance," he says. "The seed needs to be in contact with water to sprout. In tissue culture, you need to sterilize it and wash it before placing it in the growing medium. However, it must be immersed in water for 18–24 hours to break dormancy. Most protocols mention washing the seed in various solutions, but they often fail to mention the need for water immersion first. If the seed is placed in water, even for two hours, the sprout starts to emerge. At this point, even one drop of the alcohol used for sterilization will kill the seed. This crucial step is frequently omitted in many protocols. Imagine buying seeds worth hundreds of dollars and following one of these protocols for a tissue culture experiment. You'd lose all the seeds and the money you spent on them."
Another common issue involves pH levels. "Normally, in tissue culture, you set the pH at 5.75. But for cannabis, it should be a bit higher. Only a few papers mention this; most simply state 5.75. If you use that pH, everything will die after a certain time."
After extensive research and trial and error, Reza is finally reaching conclusions in his cannabis tissue culture experiments. "I eventually found the right hormones, concentrations, and medium. However, cannabis tissue culture is still in its infancy, and as we progress, there will be many other aspects to focus on. As a scientist and former academic, I'm fully aware of the importance of publishing papers. However, science is about conducting experiments that can be replicated with the same results every time, and those active in this field should adhere to that principle."