Efficient propagation of suspended HL-60 cells in a disposable bioreactor supporting wave-induced agitation at various Reynolds number

Kamil Wierzchowski,Iwona Grabowska,Maciej Pilarek

doi:10.1007/s00449-020-02386-6

Kamil Wierzchowski, Iwona Grabowska + Show 1 more

Open Access

https://doi.org/10.1007/s00449-020-02386-6

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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 ReL (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.

Highlights

The modern biopharmaceutical industry produces and commercially offers a range of animal-based bioproducts obtained through in vitro bioprocessing of mammalian and human cells, which can roughly be divided into two basic morphological categories: anchorage-dependent cells and free-floating ones
The values of R eL, as well as kLa, have been calculated for each set of operational parameters (i.e., α and ω) defining hydrodynamic conditions of wave-induced agitation during cultures of HL-60 cells performed in WAVE 25 bioreactor
It resulted in the following simplified form of dimensionless correlation applied to predict the kLa coefficients in the WAVE 25 bioreactor operated at QG = 0.5 L min−1 with the liquid phase incubated at 37 °C: kLa = 8.57 × 10−2Re0L.522 h−1

Summary

Introduction

The modern biopharmaceutical industry produces and commercially offers a range of animal-based bioproducts obtained through in vitro bioprocessing of mammalian and human cells, which can roughly be divided into two basic morphological categories: anchorage-dependent (adherent) cells and free-floating (nonadherent) ones. Instead of commonly applied rotating or tumbling stirrers used to induce fluid flow in classical bioreactor systems, continuously oscillating devices can be utilized for a gentle obtaining of homogeneous conditions in the systems for in vitro culture of the fragile animal cells. In such approaches, the agitation is achieved by 2-D horizontal oscillations of the disposable culture bag fixed in a rocker unit [6]. Efficient in vitro propagation of animal cells can be performed at the sufficient level of aeration and satisfactory values of the kLa coefficient reached at low level of R eL number characterizing wavy flow of culture medium in such single-use bioreactor platforms [9, 10]

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