A miniaturized bioreactor system for the evaluation of cell interaction with designed substrates in perfusion culture

Abstract

. (2012) AbstractIn tissue engineering, the chemical and topographical cues within three-dimensional (3D) scaffolds are normally tested using static cell cultures but applied directly to tissue cultures in perfusion bioreactors. As human cells are very sensitive to the changes of culture environment, it is essential to evaluate the performance of any chemical, and topographical cues in a perfused environment before they are applied to tissue engineering. Thus the aim of this research was to bridge the gap between static and perfusion cultures by addressing the effect of perfusion on cell cultures within 3Dscaffolds. For this we developed a scale down bioreactor system, which allows to evaluate the effectiveness of various chemical and topographical cues incorporated into our previously developed tubular ε-polycaprolactone scaffold under perfused conditions. Investigation of two exemplary cell types (fibroblasts and cortical astrocytes) using the miniaturized bioreactor indicated that: (1) quick and firm cell adhesion in 3D scaffold was critical for cell survival in perfusion culture compared with static culture, thus cell seeding procedures for static cultures might not be applicable. Therefore it was necessary to re-evaluate cell attachment on different surfaces under perfused conditions before a 3D scaffold was applied for tissue cultures, (2) continuous medium perfusion adversely influenced cell spread and survival, which could be balanced by intermittent perfusion, (3) micro-grooves still maintained its influences on cell alignment under perfused conditions, while medium perfusion demonstrated additional influence on fibroblast alignment but not on astrocyte alignment on groovedsubstrates. This research demonstrated that the mini-bioreactor system is crucial for the development 3 of functional scaffolds with suitable chemical and topographical cues by bridging the gap between static culture and perfusion culture.

The Development of a ɛ-Polycaprolactone Scaffold for Central Nervous System Repair

“…21,31 However, the significantly lower level of myelination at 28 DIV across the PCL substrates irrespective of topographical features, compared with control cultures suggested a dominant, substrate-specific effect. This effect of PCL on myelination was not due to cytotoxicity due to impurities in the PCL or degradation products 43 or the higher hydrophobicity of PCL when compared to glass coverslips.…”

“…To investigate if the topological discontinuities the micropattern, the pores, and the combined micropattern and pores in the PCL membrane 21,31 would affect CNS cell survival and myelination, we compared cell biological properties between CNS cultures on the PLL-coated flat PCL membrane (Fig. 3B), the porous PCL membrane (Fig.…”

Development of a Novel 3D Culture System for Screening Features of a Complex Implantable Device for CNS Repair

“…In this study, novel scaled down mini-chambers were developed to evaluate how cells interact with our previously fabricated tubular scaffold, with several important cell responses identified and tested. 23 It is accepted that a major problem with 3D culture systems is the survival of cells within the culture itself, with oxygen and nutrient depletion leading to a proportional necrosis of the cells, based upon their distance from the edge of the 3D system. 11 The mini-chambers, due to their configuration, also demonstrated this effect on cell survival in the absence of a porous lid, as demonstrated by the variable cell density in mini-chambers of different length, with the preference of cells to be located in areas proximal to the edges.…”

Modulation of alignment and differentiation of skeletal myoblasts by biomimetic materials

“…The bioreactor system was assembled according to T. Sun et al, 2011. 15 Briefly, the bioreactor system consisted of perfusion chambers (Fig. S1B, ESIw), a peristaltic pump (IPC-N, ISMATEC, Switzerland) (Fig.…”