Cannabis Tissue Culturing

Tissue Culturing is an interesting technique for replicating plants that has emerged over the last few years. Here we will explain what you need to know about, and how you might be using these plantlets, or so-called artificial seeds, in your own garden soon.

“The term “ tissue culture” is commonly used in a very wide sense to include in vitro aseptic culture of plant cells, tissue and organs. It is the term for “the process of growing cells artificially in the laboratory.” It involves both plant and animal cells. Tissue culture produces clones, in which all product cells have the same genotypes (unless affected by mutation during culture).” [2]

Above: Encapsulated tissue cultures of cannabis, ready-to-plant. Photo by Chimera.

Chimera offers us this thought for starters:

“Tissue culture is a whole series of practices, it can be much more complicated than just “cloning”.
Meristem culture, callus induction and regeneration, embryogenesis, organogenesis, suspension culture, microspore or anther culture, cryopreservation… I’ve been playing with TC for about 15 years. For most cultivators there is no benefit to TC as stem cuttings are easy and low-input in terms of resources, labour, etc, but it does provide some benefits in various research paradigms for germplasm storage, etc.”

Above: tissue cultures growing in nutrient solution. Photo from SkunkPharm Research.

There are some distinct advantages to using tissue cultures to propagate a garden:

~plantlets are true to the mother with no variation
~pest and disease free
~uniform growth as all plantlets are identical
~year round planting and harvesting
~new varieties can be safely introduced without contamination
~ability to scale-up in size for production of quantity with consistency of quality

Who is working on this?

Chimera, Graywolf, SkunkPharm, and almost every University around the world, as well as most large scale agribusinesses are involved in researching these methods. Fifteen years ago I saw an elaborate laboratory (solar powered) built to propagate plants for the agricultural displays at the Floriade Exposition (held every ten years in the Netherlands in a new location to showcase new technology in the plant world). There were many workstations with laminar flow hoods, rolling racks of tissues in culture, and many technicians busy behind plate glass windows for the public to see the ‘show.’

Above: A laminar flow work cabinet, photo courtesy of Chimera.

Using a laminar flow cabinet as a work bench for this technique is essential. You must prepare a sterile work environment in your laboratory.

“A laminar flow cabinet or laminar flow closet or tissue culture hood is a carefully enclosed bench designed to prevent contamination of semiconductor wafers, biological samples, or any particle sensitive materials. Air is drawn through a HEPA filter and blown in a very smooth, laminar flow towards the user. The cabinet is usually made of stainless steel with no gaps or joints where spores might collect.” [1]

Cannagraphic site member Ronbo51 wrote:

“…you would need a hood to have consistent results…You need it to do transfers and you can store plates and sterilized equipment in it after you take it out of your pressure vessel, which is the second piece you have to buy. I had a large autoclave for bulk sterilizing but also had several large pressure cookers for other things. You need to take stuff out of your pressure vessel directly to cool under the hood. Agar is pretty much sugar and even one bacteria will contaminate your plate and ruin everything…I had a clean room, practiced clean room techniques, did everything I could to keep the filthy world out of my lab but still had random contaminates. To me the biggest advantage of tissue culture would be combined with long term storage so as to insure that your genetic library remains intact and viable…”