Abstract
BackgroundOlfactory ensheathing cell (OEC) transplantation is emerging as a promising therapy for spinal cord injuries. However, outcomes are inconsistent, and the method needs improvement. Currently, cells are injected into the injury site as a suspension, and often fail to form a three-dimensional (3D) network crucial for both survival of the transplanted cells, and for regeneration of severed axons. 3D culture systems are therefore likely to improve the method. Of the many 3D culture systems available, the spheroid-producing naked liquid marble (NLM) technique is particularly advantageous compared to other platforms as it rapidly generates cell spheroids which can easily be extracted for further handling. To improve production of the spheroids, we designed and tested a device which allows fine control over vibrational stimuli to liquid marble cell cultures. We applied vibrational frequencies of 20, 60, and 80 Hz with consistent amplitude to NLM containing OECs and assessed the size and number of the 3D cell spheroids generated as well as the migratory capacity of cells cultured in the vibrated spheroids.ResultsVibrating the NLMs led to fewer and dramatically larger spheroids in comparison to non-vibrated NLMs. Of the frequencies tested, 60 Hz caused over 70-fold increase in spheroid volume. When transferred to a culture plate, the larger spheroids retained their structure after 72 h in culture, and cells that migrated out of the spheroids covered a significantly larger area compared to cells migrating out of spheroids formed at all the other frequencies tested.ConclusionsWe have shown that vibration can be used to regulate the formation of cell spheroids in NLM cultures. The ability to modulate the size of spheroids is useful for a range of 3D cell culture models and for preparing cells for in vivo transplantation.
Highlights
Olfactory ensheathing cell (OEC) transplantation is emerging as a promising therapy for spinal cord injuries
We have developed a naked liquid marble (NLM) platform (Australian provisional patent 2,017,904,456, [23]) in which droplets of cell culture medium are incubated on a superhydrophobic coating
Calibration and amplitude distribution We hypothesised that vibration can be used to modify the properties of NLM cell cultures
Summary
Olfactory ensheathing cell (OEC) transplantation is emerging as a promising therapy for spinal cord injuries. OECs require cell-cell contacts for many functions, in particular migration [18], and the lack of cell-cell contact in cell suspensions may be contributing to the low survival rate Another problem that is hampering optimal development of OEC transplantation therapies is that many of the biological functions of these cells remain unknown. Spheroid formation inside sessile LMs is highly variable, with a tendency for the cells to form non-uniform spheroid morphologies of various sizes, often with necrotic cores [20], and cell degradation at the edge of the aggregation [21] This is due to the deformation of LMs when cultured on a hard surface. Sessile marbles have a tendency to evaporate [22], causing wrinkling which further hinders efficient spheroid production
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