Culture medium and plant growth regulator regimes influence the transcriptome and cannabisin accumulation in Cannabis sativa cell suspension cultures.

Thiry M.; Sergeant K.; Legay S.; Iken M.; Renaut J.; Lutts S.; Guerriero G. · Frontiers in plant science · 2026

**Background & Methods** This study investigated how two culture media formulations (Murashige & Skoog [MS] and Linsmaier & Skoog [LS]-based) influence growth, gene expression, and cannabinoid production in Cannabis sativa cell suspension cultures derived from a non-commercial genotype (Line 43/178). Cultures were analyzed at five time points across a 14-day subculture cycle using transcriptomic analysis and untargeted metabolomics. **Key Findings** • Both MS and LS media supported comparable growth kinetics with peak biomass production reaching ≥13 g/L dry weight; however, LS medium preferentially induced expression of genes involved in hydroxycinnamic acid amide and lignanamide biosynthesis, resulting in increased accumulation of cannabisin D and cannabisin F, with marked differences in cell morphology observed between media. • Transcriptomic analysis identified 2,801 differentially expressed contigs with absolute maximum fold changes >10, revealing distinct growth-stage-dependent gene expression patterns: early activation of cell cycle and hormone-related genes, intermediate induction of stress and cell wall remodeling pathways, and late-stage senescence and redox-related processes. • LS medium-cultured cells demonstrated medium-dependent differences in total phenolic content and antioxidant activity compared to MS medium, indicating that culture medium composition critically regulates the balance between growth and secondary metabolism in Cannabis sativa cell factories. **Dosage & Administration** Not reported. **Safety & Adverse Effects** Not reported. **Evidence Quality** This is a basic research study (in vitro cell suspension culture model) rather than a clinical investigation, limiting direct applicability to human therapeutic use. The study employed a single non-commercial cannabis genotype, which restricts generalizability across cannabis varieties. While the transcriptomic and metabolomic approaches are robust, the work is observational without intervention controls beyond medium composition comparison. The study provides valuable mechanistic insights into optimizing secondary metabolite production in plant cell factories but does not assess safety, efficacy, or bioavailability in biological systems. Evidence quality is moderate for understanding culture optimization but low for clinical relevance.

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