Plant polysaccharides can exert immunomodulatory activities. In this study we provided chemical characterization of wheat cell culture-derived polysaccharides (WCCPS) and assessed their capacity to modulate inflammatory responses in mouse macrophages. The total sample (T-010) contained arabinogalactans, arabinans, glucans and xyloglucans. Fractionation by anion-exchange chromatography rendered a bound acidic fraction (B-010) and an unbound neutral fraction (UB-010). The B-010 fraction was enriched in arabinogalactans and arabinans, with some galactans, homogalacturonans, and arabinoxylans. The neutral UB-010 fraction was composed of glucans and xyloglucans. None of the WCCPS preparations triggered cytokine production on their own, but each potentiated different macrophage responses to bacterial lipopolysaccharide (LPS). The total WCCPS in T-010 increased LPS-induced tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6 secretion, whereas the acidic arabinogalactan-rich fraction B-010 boosted IL-6 release and selectively upregulated nitric oxide synthase 2 (Nos2) and cholesterol 25-hydroxylase (Ch25h) expression in response to LPS. In contrast, the neutral UB-010 fraction enhanced IL-6 levels and induced Nos2 expression without altering Ch25h expression. These results suggest that WCCPS can modulate distinct aspects of the inflammatory response, with their effects shaped by their composition and structural features. Future research will focus on elucidating the molecular mechanisms underlying the immunomodulatory activity of WCCPS.

Natural polysaccharides (PSs) have shown inhibitory effects on differentiated cancer cells (DCCs), but their activity against cancer stem cells (CSCs) remains poorly understood. Here, we report that PSs from wheat cell cultures (WCCPSs) inhibit the proliferation of both DCCs and CSCs derived from HCT-116 colorectal cancer cells. Among them, NA and DC fractions showed the strongest anti-CSC activity. NA, rich in xylose, was effective at lower concentrations, while DC, enriched in xylose and galacturonic acid (GalUA), exhibited higher potency, with a lower IC50 and preferential activity against CSCs at higher doses. WCCPSs reduced β-catenin levels, and some fractions also downregulated Ep-CAM, CD44, and c-Myc. Notably, DC increased caspase-3 without inducing cytochrome C and caspase-8 overexpression, suggesting a mechanism promoting CSC differentiation rather than apoptosis. Correlation analysis linked xylose content to reduced c-Myc expression, and GalUA levels to increased caspase-3. These results suggest that WCCPS bioactivity may be related to their monosaccharide composition. Overall, our findings support the potential of wheat-derived PSs as CSC-targeting agents that suppress self-renewal and promote differentiation, offering a promising approach to reduce tumor aggressiveness and recurrence.

There is a global need to discover effective anti-cancerous compounds from natural sources. Cultivated wheat cells can be a valuable source of non-toxic or low toxic plant-derived polysaccharides. In this study, we evaluated the anti-cancer ability of seven fractions of wheat cell culture polysaccharides (WCCPSs) in the HCT-116 colon cancer cell line. Almost all (6/7) fractions had an inhibitory effect on the proliferation of colon cancer cells, and two fractions (A-b and A-f) had considerable therapeutic indexes. The WCCPS fractions induced cell cycle arrest in the G1 phase and induced different rates of apoptosis (≤48%). Transmission and scanning electron microscopy revealed that WCCPS fractions caused apoptotic changes in the nucleus and cytoplasm, including damage to mitochondria and external morphological signs of apoptosis. In addition, the WCCPSs induced an increase in the levels of Bax, cytochrome c, and caspases 8 and 3, indicating that cell death progressed through intrinsic and extrinsic pathways of apoptosis. Furthermore, some fractions caused a significant decrease of c-Myc, b-catenin, NFkB2, and HCAM (CD 44) levels, indicating enhanced cell differentiation. Thus, for the first time, our results provide a proof of concept of the anti-cancer capacity of WCCPS fractions in colorectal cancer.

Plant polysaccharides with technical and physiologic traits attract researchers by their high physiological activity in regulation of the growth, development and protective reactions. Cell cultures allow to regulate chemical composition of synthesized substances by changing media composition and are widely used to enhance or change the biosynthesis of metabolites. The aim of this study was to investigate the influence of phytohormones 2,4-dichlorphenoxyacetic acid (2,4 –D) and abscisic acid (ABA) of culture medium on chemical composition of polysaccharides (PS), extracted from cells and extracellular liquid of wheat suspension culture. It was shown for the medium with ABA that monosaccharide composition of extracellular PS mainly represented by glucose (87%), whereas PS isolated from cells were rich for xylose and glucuronic acid. Monosaccharide composition of extracellular PS from media with 2,4-D showed 6-fold increase of arabinose, 8-fold ‒ of galactose, 5-fold ‒ of xylose and glucuronic acid, compared to extracellular PS from ABA medium. Composition of cellular PS from media with 2,4-D were mainly similar to ABA and differed by the increased amount of mannose (3-fold), and galacturonic acid (2,5-fold). Thus, regulative effect of the use of two different types of phytohormones was demonstrated on the biosynthesis of variously composed polysaccharides.

. Oligosaccharides, being a part of plant cell wall, can be secreted to the intercellular space and perform «signaling» activity on plant tissues – high physiological activity in very low concentrations. Oligosaccharide (OS) fraction was obtained from extracellular liquid wheat cell culture by size exclusion chromatography. Monosaccharide composition of extracellular OS has been determined by HPLC Ionic chromatography. It was shown that extracellular OS contain glucose, xylose, arabinose, galactose, fructose and rhamnose monosaccharide residues.

Polysaccharides were extracted from wheat suspension culture and cultivated cells. Gaz-liquid chromatography of wheat suspension culture allowed identifying the composition of polysaccharides that were found to contain arabinose, xylose, galactose, glucose, and a little amount of mannose and rhamnose. In dried cultivated cells the polysaccharide structure consisted mainly from arabinose, xylose, and galactose. It was found that the percentage of the amount of monosaccharides was positively correlated with the concentration of 2,4-D at the suspension media.

Polysaccharides (PS) from wheat cell culture were isolated by liquid-liquid extraction. The molecular mass distribution was determined by gel-permeation chromatography (GPC) using dual detectors for the simultaneous detection. It was supposed that PS sample from wheat cell culture has molecular weight of 1632 Da. The physic-chemical properties of PS such as solubility in different solvents, surface activity, ξ-potential, the pH value, polydispersity (PDI) were determined. The PS sample was soluble in water and insoluble in ethanol, acetone and chloroform. ξ-potential of PS was evaluated in order to determine its charge at different pH value from 3 to 9. As a result, the ξ values for the PS solution were negative throughout the pH range studied, varying from -2.85 mV (pH 3.0) to -21.1 (pH 9). Using tensiometry method, surface tension of the PS at the liquid/air interface was investigated. At 0.05% concentration interfacial tension decreases slowly and reaches an equilibrium value after ~ 8-8.5 hours. The pH was equal to 5.6±0.05. For a PS solution of 0.001% at pH 5.5 PDI was equal to 0.595.