Fully synthetic, biochemically inert and water-immiscible liquid perfluorochemicals (PFCs) are recognised as flexible liquid carriers/scavengers of gaseous compounds (respiratory gases mainly, i.e. O2 and CO2) and increasingly applied in bioprocess engineering. A range of unmatched physicochemical properties of liquid PFCs, i.e. outstanding chemo- and thermostability, extremely low surface tension, simultaneous hydro- and lipophobicity, which result from carbon chain substitution with fluorine atoms (the most electronegative chemical element) and the presence of intramolecular C-F bonds (the strongest single bond known in organic chemistry) have been described in detail. Exceptional propensity to solubility of respiratory gases in liquid perfluorinated compounds has been widely discussed. Advantages and disadvantages of bioprocess applications of liquid PFCs in the form of a pure PFC as well as in an emulsified form have been pointed out. A liquid PFC-mediated mass transfer intensification in various types of microbial, plant cell and animal cell culture systems: from miniaturised microlitre-scale cultures, via biomaterial-based scaffolds containing culture systems, to litre-scale bioreactors, has been reviewed and elaborated on bearing in mind the benefits of bioprocesses.
Typically applied static (i.e. non-agitated) cultures do not provide sufficient conditions for efficient propagation of suspended non-adherent cells, in general. Feasibility of small-scale wave-type agitated single-use bioreactors for gentle agitation underlies applicability of such systems for scaling-up of fragile biomass of animal cells. The basic aim of the study was to compare the results of non-adherent HL-60 cell propagation performed referentially as the batch culture in typical static (i.e. non-agitated) disposable culture flasks (50 cm3 of culture medium) and in ReadyToProcess WAVETM25 bioreactor system (GE Healthcare) equipped with disposable culture bag (300 cm3 of culture medium) subjected to continuous wave-type agitation. The density and viability of HL-60 cells were significantly higher for the bioprocess subjected to wave-type agitation, than in the reference static culture. The values of the specific rate of glucose consumption per cell (rglc=cell) exhibited by HL-60 cells maintained in the system with continuous wave-type agitation was significantly lower (i.e. up to more than 42%) than the values noted for the static culture, for exactly the same time-points of two compared cultures. The results of the studies undoubtedly and comprehensively confirmed the applicability of the studied disposable bioreactor with wave-induced agitation as the right platform for proceeding the propagation of non- adherent HL-60 cells and for providing the culture conditions required by HL-60 cells for sustainable metabolism.
The aim of this study was to determine the solubility of CO2 in perfluorodecalin (PFD) which is frequently used as efficient liquid carrier of respiratory gases in bioprocess engineering. The application of perfluorinated liquid in a microsystem has been presented. Gas-liquid mass transfer during Taylor (slug) flow in a microchannel of circular cross section 0.4 mm in diameter has been investigated. A physicochemical system of the absorption of CO2 from the CO2/N2 mixture in perfluorodecalin has been applied. The Henry’s law constants have been found according to two theoretical approaches: physical (H = 1.22·10-3 mol/m3Pa) or chemical (H = 1.26·10-3 mol/m3Pa) absorption. We are hypothesising that the gas-liquid microchannel system is applicable to determine the solubility of respiratory gases in perfluorinated liquids.
A mathematical model of a hybrid culture system supported with a stationary layer of liquid perfluorochemical (PFC) as a source of O2 for cells which grow in the aqueous phase of culture medium has been developed and discussed. The two-substrate Monod kinetics without inhibition effects, i.e. the Tsao-Hanson equation, has been assumed to characterise the biomass growth. The Damköhler number which relates the growth rate to the mass transfer effects has been used to appraise the regime (i.e. diffusion-limited or kinetics) of the whole process. The proposed model predicted accurately previously published data on the submerged batch cultures of Nicotiana tabacum BY-2 heterotrophic cells performed in a culture system supported with a stationary layer of hydrophobic perfluorodecalin as a liquid O2 carrier. Estimated values of the parameters of the model showed that the process proceeded in the kinetics regime and the growth kinetics, not the effects of the mass transfer between aqueous phase and liquid PFC, had essential influence on the growth of biomass.
Kinetic resolution of (R)- and (S)-mandelic acid by its transesterification with vinyl acetate catalysed by Burholderia cepacia lipase has been studied. The influence of the initial substrate concentration on the kinetics of process has been investigated. A modified ping-pong bi-bi model of enzymatic transesterification of (S)-mandelic acid including substrate inhibition has been developed. The values of kinetic parameters of the model have been estimated. We have shown that the inhibition effect revealed over a certain threshold limit value of the initial concentration of substrate.
One of the actual challenges in tissue engineering applications is to efficiently produce as high of number of cells as it is only possible, in the shortest time. In static cultures, the production of animal cell biomass in integrated forms (i.e. aggregates, inoculated scaffolds) is limited due to inefficient diffusion of culture medium components observed in such non-mixed culture systems, especially in the case of cell-inoculated fiber-based dense 3D scaffolds, inside which the intensification of mass transfer is particularly important. The applicability of a prototyped, small-scale, continuously wave-induced agitated system for intensification of anchorage-dependent CP5 chondrocytes proliferation outside and inside three-dimensional poly(lactic acid) (PLA) scaffolds has been discussed. Fibrous PLA-based constructs have been inoculated with CP5 cells and then maintained in two independent incubation systems: (i) non-agitated conditions and (ii) culture with wave-induced agitation. Significantly higher values of the volumetric glucose consumption rate have been noted for the system with the wave-induced agitation. The advantage of the presented wave-induced agitation culture system has been confirmed by lower activity of lactate dehydrogenase (LDH) released from the cells in the samples of culture medium harvested from the agitated cultures, in contrast to rather high values of LDH activity measured for static conditions. Results of the proceeded experiments and their analysis clearly exhibited the feasibility of the culture system supported with continuously wave-induced agitation for robust proliferation of the CP5 chondrocytes on PLA-based structures. Aside from the practicability of the prototyped system, we believe that it could also be applied as a standard method offering advantages for all types of the daily routine laboratory-scale animal cell cultures utilizing various fiber-based biomaterials, with the use of only regular laboratory devices.