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

Abstract

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.

Keywords: COVID-19; GvHD; Mesenchymal stromal cells; Supply chain.

Fig. 1

Experimental design. To simulate the different shipment modalities, cells stored in LN2 vapour were thawed directly at 37 °C or after incubation for 18 h in dry ice. Cell viability and apoptosis were determined immediately after thawing. Immunophenotype, immunomodulation, and apoptosis were analysed after one week in culture. Population doubling was determined by counting cells at days 0, 2, 5 and 7. CFU-F potential was determined on cells plated immediately after thawing or plated after one week in culture (n = 3)

Fig. 2

Analyses of cell viability, apoptosis rates and population doubling. a Histograms of the percentage of viable cells after thawing, measured with trypan blue exclusion assay. b Population doubling (PD) calculated at Days 2, 5 and 7 of culture post-thawing. c Representative cytofluorimetric plots of apoptosis analyses and relative results expressed as percentages. Viability (V), early apoptosis (EA), late apoptosis (LA), and necrosis (N) of cells stored in LN₂ and dry ice were analysed immediately after cell thawing (Day 0) or after one week in culture (Day 7). Data are expressed as mean ± SD of three independent experiments. Statistical analysis was performed with GraphPad Prism software and Student’s t-test did not evidence any statistical difference

Fig. 3

UC-MSCs immunomodulation properties. The anti-proliferative activity of UC-MSCs was analysed by flow cytometry after PBMC labelling with CFSE. Statistical significance is relative to the proliferation of activated lymphocytes in the absence of MSCs (n = 3) (*p < 0.05; ** p < 0.01)

Fig. 4

UC-MSC clonogenic potential. a Representative images of CFU-F stained with crystal violet. b Histograms showing the absolute number of CFU-F obtained from cells thawed after storage in dry ice compared to control cells, thawed after storage in LN₂. No significant differences were detected