Logistics of an advanced therapy medicinal product during COVID-19 pandemic in Italy: successful delivery of mesenchymal stromal cells in dry ice.

BACKGROUND: During the coronavirus disease-2019 (COVID-19) pandemic, Italian hospitals faced the most daunting challenges of their recent history, and only essential therapeutic interventions were feasible. From March to April 2020, the Laboratory of Advanced Cellular Therapies (Vicenza, Italy) received requests to treat a patient with severe COVID-19 and a patient with acute graft-versus-host disease with umbilical cord-derived mesenchymal stromal cells (UC-MSCs). Access to clinics was restricted due to the risk of contagion. Transport of UC-MSCs in liquid nitrogen was unmanageable, leaving shipment in dry ice as the only option. METHODS: We assessed effects of the transition from liquid nitrogen to dry ice on cell viability; apoptosis; phenotype; proliferation; immunomodulation; and clonogenesis; and validated dry ice-based transport of UC-MSCs to clinics. RESULTS: Our results showed no differences in cell functionality related to the two storage conditions, and demonstrated the preservation of immunomodulatory and clonogenic potentials in dry ice. UC-MSCs were successfully delivered to points-of-care, enabling favourable clinical outcomes. CONCLUSIONS: This experience underscores the flexibility of a public cell factory in its adaptation of the logistics of an advanced therapy medicinal product during a public health crisis. Alternative supply chains should be evaluated for other cell products to guarantee delivery during catastrophes.

Multiple time scales in multi-state models.

In multi-state models, it has been the tradition to model all transition intensities on one time scale, usually the time since entry into the study ('clock-forward' approach). The effect of time since an intermediate event has been accommodated either by changing the time scale to time since entry to the new state ('clock-back' approach) or by including the time at entry to the new state as a covariate. In this paper, we argue that the choice of time scale for the various transitions in a multi-state model should be dealt with as an empirical question, as also the question of whether a single time scale is sufficient. We illustrate that these questions are best addressed by using parametric models for the transition rates, as opposed to the traditional Cox-model-based approaches. Specific advantages are that dependence of failure rates on multiple time scales can be made explicit and described in informative graphical displays. Using a single common time scale for all transitions greatly facilitates computations of probabilities of being in a particular state at a given time, because the machinery from the theory of Markov chains can be applied. However, a realistic model for transition rates is preferable, especially when the focus is not on prediction of final outcomes from start but on the analysis of instantaneous risk or on dynamic prediction. We illustrate the various approaches using a data set from stem cell transplant in leukemia and provide supplementary online material in R.

Assessment of viability and osteogenic ability of human mesenchymal stem cells after being stored in suspension for clinical transplantation.

Human mesenchymal stem cells (MSCs) were suspended in phosphate-buffered saline (PBS) and stored up to 24 h at 4 degrees C, 24 degrees C, and 37 degrees C. More than 80% viability was maintained at any temperature for at least 1 h, then gradually decreased over time. After 24 h, the viabilities at 4 degrees C, 24 degrees C, and 37 degrees C were about 81%, 70%, and 62%, respectively. The MSCs suspended/stored in PBS at 4 degrees C for 24 h also exhibited in vitro osteogenic differentiation capability as evidenced by mineralized matrix formation as well as high alkaline phosphatase activity when cultured in an osteogenic medium. Furthermore, in vivo implantation experiments using the MSCs also demonstrated new bone formation. Because MSCs are known to possess multipotential stem cell characteristics, these data indicate that human MSCs stored in PBS at 4 degrees C could be delivered to distant medical facilities for the purpose of hard tissue and other types of tissue regeneration therapy.