Banking of AT-MSC and its Influence on Their Application to Clinical Procedures

Abstract

Processing of MSCs to obtain a therapeutic product consists of two main steps: 1) the in vitro expansion of the cells until an appropriate number of them is obtained, and 2) freezing and storage of the expanded cells. The last step is critical and must be optimized so that after thawing the cells retain all their physiological properties including the secretory function. In this paper, we evaluated physiological parameters of AT-MSC’s after a full cycle of their processing, particularly freezing and storing at the liquid nitrogen vapor temperature. Based on the recovered proliferative and secretory capacities of the thawed cells, we have designed the optimal technique for processing of MSCs for clinical applications. In our work, we tried to select the best DMSO-based cryoprotectant mixture on the base of post thawing fully retain their properties. We have demonstrated the effectiveness of the use of DMSO in various configurations of the constituent cryoprotective fluids. We have also shown that AT-MSCs that show control levels in most standard tests (viability, shape, culture behaviour, and proliferative properties) after thawing, may show transient variations in some important physiological properties, such as the level of secreted growth factors. Obtained results let us to indicate how to optimize the AT-MSC preparation process for clinical applications. We suggest that before their clinical application the cells should be cultured for at least one passage to recover their physiological stability and thus assure their optimal therapeutic potential.

Optimization of Mesenchymal Stromal Cell (MSC) Manufacturing Processes for a Better Therapeutic Outcome

“…Different methods, rates of cooling and compositions of cryoprotectants have been developed ( 84 , 85 , 87 ). The most widely-used cryoprotectant to date is dimethyl sulfoxide (DMSO), although there are different excipient formulations that can give better performances in post-thaw viability ( 88 ). 10% DMSO could be supplemented with a buffer containing reagents ranging from 5% Human Albumin, Human Serum or Human Plasma A/B to more complex formulations involving Dextran-40, Lactobionate, Sucrose, Mannitol, Glucose, Adenosine or Glutathione ( 89 ).…”

“…Once the MSC has been thawed, the final characterization and delivery to the patient must be performed. Post-thawing release criteria should include parameters such as viability, recovery, phenotyping and potency assay ( 87 , 88 ). In our experience, thawing of cryopreserved cells is a critical step, it must be done quickly.…”

A Comparative Study of the Bone Marrow- and Umbilical Cord-Derived Mesenchymal Stem Cells (MSCs) Efficiency on Generating MSC-Educated Macrophages (MEMs)

“…In many studies, MEMs are usually educated by MSCs derived from BM, however, since the collection process of MSCs from BM has some limitations such as invasive acquisition procedure, higher risk of infectious diseases transmission, donor's age, and limited proliferative potential of MSCs (Pittenger et al, 2019), exploiting alternative sources could be beneficial. UC is a prenatal organ which composed of various anatomical parts like the amniotic membrane (UCM), perivascular region (PRV), and the central part of UC constituted by Wharton's Jelly (WJ) (Semenova et al, 2021). MSCs can be rapidly isolated from UC without any risk or discomfort for the donor.…”