Development of a human umbilical cord-derived mesenchymal stromal cell-based advanced therapy medicinal product to treat immune and/or inflammatory diseases

Abstract

Background Umbilical cord-derived mesenchymal stromal cells (UC-MSCs) revealed their key role in immune regulation, offering promising therapeutic perspectives for immune and inflammatory diseases. We aimed to develop a production process of an UC-MSC-based product and then to characterize UC-MSC properties and immunomodulatory activities in vitro, related to their clinical use and finally, to transfer this technology to a good manufacturing practice (GMP) compliant facility, to manufacture an advanced therapy medicinal product (ATMP). Methods Fifteen human umbilical cords (UCs) were collected to develop the production process. Three batches of UC-MSCs from a single donor were characterized at basal state and after in vitro pro-inflammatory stimulation by interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα). Proliferation, immunophenotype, activation markers’ expression and the inhibition of T cell proliferation were assessed. Finally, this technology was transferred to a GMP-compliant facility to manufacture an UC-MSC-based ATMP, from a single donor, using the explant method followed by the establishment of master and work cell stocks. Results Twelve UCs were processed successfully allowing to isolate UC-MSCs with doubling time and population doubling remaining stable until passage 4. CD90, CD105, CD73, CD44, CD29, CD166 expression was positive; CD14, CD45, CD31, HLA-DR, CD40, CD80 and CD86 expression was negative, while CD146 and HLA-ABC expression was heterogeneous. Cell morphology, proliferation and immunophenotype were not modified by inflammatory treatment. Indoleamine 2,3-dioxygenase (IDO) expression was significantly induced by IFNγ and IFNγ + TNFα versus non-treated cells. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) expression was induced significantly after priming. T cell proliferation was significantly decreased in the presence of UC-MSCs in a dose-dependent manner. This inhibitory effect was improved by IFNγ or IFNγ + TNFα, at UC-MSCs:PBMC ratio 1:10 and 1:30, whereas only IFNγ allowed to decrease significantly T cell proliferation at ratio 1:100. The manufacturing process of the UC-MSC-based ATMP was qualified and authorized by the French regulatory agency for clinical use (NCT04333368). Conclusion This work allowed to develop an investigational UC-MSC-based ATMP authorized for clinical use. Our results showed that an inflammatory environment preserves the biological properties of UC-MSCs with an improvement of their immunomodulatory functions.

Treatment of COVID-19-associated ARDS with mesenchymal stromal cells: a multicenter randomized double-blind trial

“…Our double-blind, randomized, placebo-controlled trial has several other key strengths, including serial determinations of a wide array of inflammation and immunity-related biomarkers at several times until D14, monitoring of allo-immunization, and thorough characterization of the final UC-MSC-based product. Instead of assessing the three-lineage differentiation of our cells, we rather used a potency assay more specific for their intended immunomodulatory effect and to this end performed mixed lymphocyte reactions which confirmed their inhibitory effect on allogeneic T lymphocytes [ 34 ]. A final key strength is the multicenter design.…”

Large-Scale Expansion of Human Umbilical Cord-Derived Mesenchymal Stem Cells in a Stirred Suspension Bioreactor Enabled by Computational Fluid Dynamics Modeling

“…While for the batch culture, the cells stopped proliferating on day 5 and followed a declined cell density and viability ( Figure 6 B,C). Notably, the maximum cell density of fed-batch culture (7.3 × 10 5 cells/mL on day 7) is 2.9 folds of that of batch culture, which could achieve 20.2 times expansion (compared to seeded cells), significantly higher than 2D plates culture (12.0 × 10 5 cells/mL, 8.9 folds) and most of the reported work [ 41 , 42 , 43 , 44 , 45 ].…”

Banking of perinatal mesenchymal stem/stromal cells for stem cell-based personalized medicine over lifetime: Matters arising

“…Interestingly, frozen-thawed MSCs primed by interferon-gamma (IFN-γ) in the culture medium for 48 h prior to cryopreservation may partly avoid the lysis by activated T cells[ 34 ]. The mechanism of MSC-mediated immunosuppression has been previously proposed as IDO expression in BM-MSCs[ 35 ] as well as UC-MSCs[ 36 ] induced by IFN-γ. However, while pre-licensing MSCs with IFN-γ have been seen to enhance their IDO expression in vitro [ 37 , 38 ], the pre-licensed MSCs by IFN-γ, compared to the unstimulated MSCs, have let to the loss of their effectiveness in rescuing retinal ganglion cells in a retinal ischemia/reperfusion injury mouse model[ 37 ].…”