Efficient propagation of suspended HL-60 cells in a disposable bioreactor supporting wave-induced agitation at various R
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RESEARCH PAPER
Efficient propagation of suspended HL‑60 cells in a disposable bioreactor supporting wave‑induced agitation at various Reynolds number Kamil Wierzchowski1 · Iwona Grabowska2 · Maciej Pilarek1 Received: 19 March 2020 / Accepted: 29 May 2020 © The Author(s) 2020
Abstract Growth of human nonadherent HL-60 cell cultures performed in disposable bioreactor under various hydrodynamic conditions of 2-D wave-assisted agitation has been compared and discussed. Influence of Reynolds number for liquid (ReL) and the kLa coefficient, as key parameters characterized the bioprocessing of HL-60 cells in ReadyToProcess WAVETM 25 system, on reached values of the apparent maximal specific growth rate (μmax) and the specific yield of biomass (Y*X/S) has been identified. The values of R eL (i.e., 510–10,208), as well as kLa coefficient (i.e., 2.83–13.55 h−1), have been estimated for the cultures subjected to wave-induced mixing, based on simplified dimensionless correlation for various presents of WAVE 25 system. The highest values of apparent μmax = 0.038 h−1 and Y*X/S = 25.64 × 108 cells gglc−1 have been noted for cultures independently performed at wave-induced agitation characterized by ReL equaled to 5104 and 510, respectively. The presented results have high applicability potential in scale-up of bioprocesses focused on nonadherent animal cells, or in the case of any application of disposable bioreactors presenting similitude. Keywords Disposable (single-use) bioreactor · Wave-type agitation · Re number · kLa coefficient · Human hematopoietic HL-60 cells · Nonadherent cell propagation List of symbols AW Specific absorbency, – aLDH Activity of lactate dehydrogenase, µkat L−1 am Metabolic activity, µkat L−1 CCO2 CO2 concentration in the gas phase, % CO2 O2 concentration in the gas phase, % DL O2 diffusion coefficient in water, m2 s−1 d Dilution factor, – k Number of hemocytometer squares occupied by cells, – kLa Volumetric liquid-side mass transfer coefficient, h−1 L Length of culture bag, m ReL Reynolds number for liquid phase, – ReG Reynolds number for gas phase, – * Maciej Pilarek [email protected] 1
Faculty of Chemical and Process Engineering, Warsaw University of Technology, Waryńskiego 1, 00‑645 Warsaw, Poland
Faculty of Biology, University of Warsaw, Miecznikowa 1, 02‑096 Warsaw, Poland
2
r*glc/cell Specific glucose consumption rate, g h−1 cell−1 Sc Schmidt number for liquid phase, – T Temperature, °C QG Gas flow rate, L min−1 VL Volume of culture medium, L X Density of HL-60 cells in culture medium, cell mL−1 X0 Initial density of HL-60 cells at starting point of culture, cell mL−1 x Number of cells counted in hemocytometer, – Y*X/S Specific yield of biomass from subtract, cells gglc−1 Z Viability of the cells, % z Number of alive cells, Greek symbols α Angle of oscillations, ° ΔA Absorbency change, – ΔCglc Change of glucose concentration, g L−1 Δt Time interval, h µmax Apparent maximal specific growth rate, s−1 νL Liquid phase kinematic visco
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