Application of a simple unstructured kinetic and cost of goods models to support T-cell therapy manufacture.

Manufacturing of cell therapy products requires sufficient understanding of the cell culture variables and associated mechanisms for adequate control and risk analysis. The aim of this study was to apply an unstructured ordinary differential equation-based model for prediction of T-cell bioprocess outcomes as a function of process input parameters. A series of models were developed to represent the growth of T-cells as a function of time, culture volumes, cell densities, and glucose concentration using data from the Ambr®15 stirred bioreactor system. The models were sufficiently representative of the process to predict the glucose and volume provision required to maintain cell growth rate and quantitatively defined the relationship between glucose concentration, cell growth rate, and glucose utilization rate. The models demonstrated that although glucose is a limiting factor in batch supplied medium, a delivery rate of glucose at significantly less than the maximal specific consumption rate (0.05 mg 1 × 106  cell h-1 ) will adequately sustain cell growth due to a lower glucose Monod constant determining glucose consumption rate relative to the glucose Monod constant determining cell growth rate. The resultant volume and exchange requirements were used as inputs to an operational BioSolve cost model to suggest a cost-effective T-cell manufacturing process with minimum cost of goods per million cells produced and optimal volumetric productivity in a manufacturing settings. These findings highlight the potential of a simple unstructured model of T-cell growth in a stirred tank system to provide a framework for control and optimization of bioprocesses for manufacture.

Using novel methodologies to examine the impact of artificial light at night on the cortisol stress response in dispersing Atlantic salmon (Salmo salar L.) fry.

Artificial light at night (ALAN) is gaining recognition as having an important anthropogenic impact on the environment, yet the behavioural and physiological impacts of this stressor are largely unknown. This dearth of information is particularly true for freshwater ecosystems, which are already heavily impacted by anthropogenic pressures. Atlantic salmon (Salmo salar L.) is a species of conservation and economic importance whose ecology and behaviour is well studied, making it an ideal model species. Recent investigations have demonstrated that salmon show disrupted behaviour in response to artificial light; however, it is not yet clear which physiological processes are behind the observed behavioural modifications. Here, two novel non-invasive sampling methods were used to examine the cortisol stress response of dispersing salmon fry under different artificial lighting intensities. Fish egg and embryos were reared under differing ALAN intensities and individual measures of stress were subsequently taken from dispersing fry using static sampling, whereas population-level measures were achieved using deployed passive samplers. Dispersing fry exposed to experimental confinement showed elevated cortisol levels, indicating the capacity to mount a stress response at this early stage in ontogenesis. However, only one of the two methods for sampling cortisol used in this study indicated that ALAN may act as a stressor to dispersing salmon fry. As such, a cortisol-mediated response to light was not strongly supported. Furthermore, the efficacy of the two non-invasive methodologies used in this study is, subject to further validation, indicative of them proving useful in future ecological studies.

Automated, scalable culture of human embryonic stem cells in feeder-free conditions.

Large-scale manufacture of human embryonic stem cells (hESCs) is prerequisite to their widespread use in biomedical applications. However, current hESC culture strategies are labor-intensive and employ highly variable processes, presenting challenges for scaled production and commercial development. Here we demonstrate that passaging of the hESC lines, HUES7, and NOTT1, with trypsin in feeder-free conditions, is compatible with complete automation on the CompacT SelecT, a commercially available and industrially relevant robotic platform. Pluripotency was successfully retained, as evidenced by consistent proliferation during serial passage, expression of stem cell markers (OCT4, NANOG, TRA1-81, and SSEA-4), stable karyotype, and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Automation of hESC culture will expedite cell-use in clinical, scientific, and industrial applications.