Mesenchymal stem cells (MSCs) are widely used in cell therapy due to their convenience, multiline differentiation potential, reproducible protocols, and biological properties. The potential of MSCs to impregnate magnetic microcapsules and their possible influence on cell function and ability to response to magnetic field have been explored. Interestingly, the cells suspended in media show much higher ability in internalization of microcapsules, then MSCs adhere into the surface. There is no significant effect of microcapsules on cell toxicity compared with other cell line-capsule internalization reported in literature. Due to internalization of magnetic capsules by the cells, such cell engineering platform is responsive to external magnetic field, which allows to manipulate MSC migration. Magnetically sorted MSCs are capable to differentiation as confirmed by their conversion to adipogenic and osteogenic cells using standard protocols. There is a minor effect of capsule internalization on cell adhesion, though MSCs are still able to form spheroid made by dozen of thousand MSCs. This work demonstrates the potential of use of microcapsule impregnated MSCs to carry internalized micron-sized vesicles and being navigated with external magnetic signaling.
Plant cell culture is traditionally viewed as a unique artifi cially created biological system representing a heterogenous population of dedifferentiated cells. This system undergoes a continuous process of autoselection based on the intensity and stability of cell proliferation. We discuss here the details of formation and regulation of isoprenoid biosynthesis in plant cell in vitro based on literature survey and our research. Obviously, secondary metabolism differs in cell culture compared to the plant per se , because in cell culture metabolites are synthesized and compartmentalized within a single heterotrophic cell with sparse or underdeveloped vacuoles and plastids. For example, in plant cell cultures isoprenoid biosynthesis via MVA pathway was found to be more active than via plastid-localized MEP pathway. Also, it was hypothesized that cell cultures preferably produce metabolites, which promote cell proliferation and growth. Indeed, cell cultures of Dioscorea deltoidea produced mainly furostanol glycosides, which promoted cell division. Triterpene glycosides (ginsenosides) in the cell cultures of various Panax species are represented mainly by Rg-and Rb-groups. Rb ginsenosides are predominantly found as malonyl-esters that may infl uence their intracellular localization.Despite the difference in the isoprenoid composition in plant and cell culture the latter became an attractive source of phytochemicals as an alternative to plant harvesting. We provide in this chapter the guidelines to biotechnological production of plant isoprenoids using plant cell cultures and discuss the optimal methods of bioreactor-based cultivation and cryopreservation of plant cell collections.
Dioscorea deltoidea is a medicinal plant valued for its high content of steroidal glycosides (SG) – bioactive compounds with cardioprotective and immunomodulation actions, also used to treat reproductive system disorders. To overcome the limitations of natural resources of this species, a suspension cell culture of D. deltoidea was developed as a renewable and ecologically sustainable source of raw biomass and SG. Cell culture demonstrated stable and intensive growth in the laboratory (20 L) and industrial (630 L) bioreactors operated under a semi-continuous regime (specific growth rate 0.11–1.12 day−1, growth index 3.5–3.7). Maximum dry weight accumulation (8.5–8.8 g/L) and SG content (47–57 mg/g DW) were recorded during the stationary phase. Bioreactor-produced cell biomass contained inorganic macro (К, Ca, Mg, Na) and micro (Zn, Mn, Fe, B, Al, Cu, Cr, Se, Co, Ni) elements in concentrations within the safe range of dietary recommendations. Acute toxicity test showed no or insignificant changes in organ weight, hematological panel and blood biochemistry of laboratory animals fed with 2000 and 5000 mg/kg dry biomass. The results suggest that cell culture of D. deltoidea grown in bioreactors has great potential to be used as functional foods and a component of specialized dietary supplements in complex therapy of reproductive system disorders and mineral deficiency.
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