Method of production of phytocannabinoids for use in medical treatments

Abstract

A method of producing cannabinoids for use in medical treatments by growing cultured Cannabis sativa plant cells through tissue culture, the method comprising the steps of: selecting Cannabis sativa leaf tissue for culture; and growing a tissue culture from the selected leaf tissue in a liquid based medium whilst controlling the light exposure of the tissue culture to control the cannabinoid content of the tissue culture. Control of the light exposure can enable the phytocannabinoid content of the grown tissue culture to be tailored to the use intended for the tissue culture. For example, the THC content of the tissue culture can be controlled to be maximised or minimised depending on the intended use. Use of tissue culture is beneficial as compared to prior art methods as it allows for genetic consistency and reduces the resources necessary to produce plant cells containing phytocannabinoids.

1 . A Cannabis cell suspension culture comprising:
 a. a liquid based plant cell culture medium; and   b. Cannabis callus cells.   
     
     
         2 . The cell culture of  claim 1 , further comprising cannabinoids, other phytoconstituents, or any combination thereof. 
     
     
         3 . The cell culture of  claim 2 , wherein the cell suspension culture is grown under controlled UV light exposure and wherein the cell suspension culture further comprises a controlled content of cannabinoids, a controlled content of other phytoconstituents, or any combination thereof. 
     
     
         4 . The cell culture of  claim 2 , wherein the concentration of cannabinoids in the culture ranges from about 1% (g/mL) to about 60% (g/mL). 
     
     
         5 . The cell culture of  claim 2 , wherein the cannabinoids comprise THC and CBD in a ratio of about 1:1 to about 10:1. 
     
     
         6 . The cell culture of  claim 2 , wherein the cell suspension culture is about 10 to about 28 days-old. 
     
     
         7 . The cell culture of  claim 2 , wherein the cell suspension culture comprises Cannabis cells at a density ranging from about 10,000 cells/mL to about 100,000 cells/mL. 
     
     
         8 . The cell culture of  claim 1 , wherein the Cannabis cell suspension culture comprises liquid cell culture medium inoculated with callus cells grown on solid cell culture media. 
     
     
         9 . The cell culture of  claim 8 , wherein the solid media cell culture is about 15 to about 45 days-old. 
     
     
         10 . The cell culture of  claim 8 , wherein the liquid cell culture medium comprises about 0.5 to about 1.5 g/mL or about 1 g/mL of the Cannabis callus cells. 
     
     
         11 . The cell culture of  claim 8 , wherein the solid media cell culture is grown under a full spectrum of photosynthetically active radiation (PAR) and wherein the solid media cell culture is grown under minimized exposure to UV radiation. 
     
     
         12 . The cell culture of  claim 11 , wherein the concentration of cannabinoids ranges from about 0.05% to about 2%. 
     
     
         13 . The cell culture of  claim 11 , wherein the density of Cannabis cells in the cell suspension culture ranges from about 10,000 to about 100,000 cells/mL. 
     
     
         14 . The cell culture of  claim 11 , wherein the solid media cell culture is about 10 to about 28 days-old. 
     
     
         15 . The cell culture of  claim 1 , wherein the Cannabis cell suspension culture comprises liquid cell culture medium inoculated with callus cells grown in liquid cell culture media. 
     
     
         16 . The cell culture of  claim 1 , wherein the cell suspension culture comprises a volume ranging from about 1 mL to about 60 mLs, from about 40 mL to about 100 mLs, or from about 90 mLs to about 3 Ls or more. 
     
     
         17 . The cell culture of  claim 1 , wherein the cell suspension culture comprises a volume ranging from about 100 Ls to about 5000 Ls or more. 
     
     
         18 . The cell culture of  claim 1 , further comprising a cell culture container or a bioreactor comprising the cell suspension culture. 
     
     
         19 . The cell culture of  claim 1 , wherein the cell culture medium comprises naphthalene acetic acid (NAA). 
     
     
         20 . The cell culture of  claim 1 , wherein the liquid cell culture medium comprises water, macronutrients, micronutrients, vitamins, amino acids or nitrogen supplements, carbon sources, organic supplements, growth regulators, or any combination thereof. 
     
     
         21 . The cell culture of  claim 20 , wherein the liquid cell culture medium comprises NAA, cannabinoids, other phytoconstituents, or any combination thereof. 
     
     
         22 . The cell culture of  claim 1 , wherein the liquid cell culture medium is Anderson rhododendron medium, CHU(N6) medium, CLC/Ipomoea medium, Chee & Pool (C2D) vitis medium, De greef & jacobs medium, DKW/JUNGLANS medium, Eriksson(er) medium, Gamborg B5 medium, Gresshof & doy (DBM2) medium, Hellers medium, kao michayluk medium, knudson corchid medium, Lindemann Orchid medium, Litvay medium, Linsmaier & Skoog medium, McCowns woody plant medium, Murashige & Skoog medium, Murashige & Miller medium, nitsch medium, NLN medium, orchimax medium, quoirin & Lepoivre medium, rugini olive medium, schenk & hildebrandt medium, S-Medium, vacin and went medium, white medium, westvaco WV3 medium, and any combination thereof, or any modification thereof. 
     
     
         23 . The cell culture of  claim 1 , wherein the Cannabis callus cells are derived from Cannabis leaves, flower buds, flower cells, bract cells, trichomes, or any combination thereof. 
     
     
         24 . The cell culture of  claim 1 , wherein the Cannabis cell suspension culture is comprised in conditions for storage. 
     
     
         25 . The cell culture of  claim 1 , wherein the Cannabis callus cells are derived from Cannabis leaves. 
     
     
         26 . The cell culture of  claim 25 , wherein the leaves are selected to provide the correct phytocannabinoid composition 
     
     
         27 . A Cannabis cell suspension culture comprising:
 a. a liquid based plant cell culture medium;   b. a cell culture container or bioreactor comprising the cell culture medium; and   c. Cannabis callus cells.   
     
     
         28 . The cell culture of  claim 27 , further comprising cannabinoids, other phytoconstituents, or any combination thereof. 
     
     
         29 . A Cannabis cell suspension culture comprising:
 a. a liquid based plant cell culture medium comprising naphthalene acetic acid (NAA);   b. Cannabis callus cells; and   c. cannabinoids, other phytoconstituents, or any combination thereof.   
     
     
         30 . The cell culture of  claim 29 , wherein the cell suspension culture is derived from a subculture of Cannabis callus cells. 
     
     
         31 . The cell culture of  claim 29 , wherein the cell suspension culture is about 10 to about 28 days-old. 
     
     
         32 . The cell culture of  claim 29 , wherein the cell suspension culture is about 15 to about 45 days-old. 
     
     
         33 . A solid media Cannabis cell culture comprising:
 a. a solid-based plant cell culture medium comprising NAA; and   b. Cannabis callus cells.   
     
     
         34 . The solid media Cannabis cell culture of  claim 33 , wherein the solid media Cannabis cell culture is about 15 to about 45 days-old. 
     
     
         35 . The solid media Cannabis cell culture of  claim 33 , wherein the solid media cell culture is grown under minimized exposure to UV radiation. 
     
     
         36 . A kit for preparing a Cannabis cell suspension culture of  claim 1 , the kit comprising:
 a. a plant cell culture medium;   b. Cannabis callus cells; and   c. optionally a cell culture container or a bioreactor.   
     
     
         37 . The kit of  claim 36 , wherein the plant cell culture medium is a liquid based culture medium, a solid culture medium, or both. 
     
     
         38 . The kit of  claim 36 , wherein the cell culture container comprises the Cannabis callus cells.