Cryopreservation of Human Mesenchymal Stem Cells in an Allogeneic Bioscaffold based on Platelet Rich Plasma and Synovial Fluid

Abstract

Transplantation of mesenchymal stem cells (MSCs) has emerged as an alternative strategy to treat knee osteoarthritis. In this context, MSCs derived from synovial fluid could provide higher chondrogenic and cartilage regeneration, presenting synovial fluid as an appropriate MSCs source. An allogeneic and biomimetic bioscaffold composed of Platelet Rich Plasma and synovial fluid that preserve and mimics the natural environment of MSCs isolated from knee has also been developed. We have optimized the cryopreservation of knee-isolated MSCs embedded within the aforementioned biomimetic scaffold, in order to create a reserve of young autologous embedded knee MSCs for future clinical applications. We have tested several cryoprotectant solutions combining dimethyl sulfoxide (DMSO), sucrose and human serum and quantifying the viability and functionality of the embedded MSCs after thawing. MSCs embedded in bioscaffolds cryopreserved with DMSO 10% or the combination of DMSO 10% and Sucrose 0,2 M displayed the best cell viabilities maintaining the multilineage differentiation potential of MSCs after thawing. In conclusion, embedded young MSCs within allogeneic biomimetic bioscaffold can be cryopreserved with the cryoprotectant solutions described in this work, allowing their future clinical use in patients with cartilage defects.

Advances in the slow freezing cryopreservation of microencapsulated cells

“…They have been associated with other function such as thermal hysteresis or interaction with membranes and/or membrane proteins, and due to these characteristics they were employed in the freezing protocols of different cell lines [40][41][42] for example, by the use of human serum. Moreover, the variation between the used human serum batches can be significant; some reports showed beneficial effects when it was included in the CPA solution [46], and others not [47]. 3B).…”

“…The only method to preserve cell-based systems or products for long periods is with the use of low temperatures. Thus, the efforts to cryopreserve cell-based products made of natural polymers (e.g., agarose, hyaluronic acid, fibrin or collagen) or synthetic polymers (e.g., PEG) have linearly grown with its reflection in more publications in last decades[22,30,47,79,81]. Alginate has been one of the most often used polysaccharides to form cellularized bioscaffolds due to its potential as sustained drug delivery or cell delivery systems for the treatment of different diseases (e.g., diabetes, hepatic diseases, neurodegenerative or cancer among others)[82][83][84][85].…”

Storage stability of liposomes stored at elevated subzero temperatures in DMSO/sucrose mixtures

“…Examples of such compounds are disaccharides (sucrose, trehalose) and polymers including ficoll, hydroxyethyl starch (HES) and polyvinyl pyrrolidone (PVP) [ 12 – 14 ]. Multiple reports exist in which such combinations were successfully employed for cryopreservation of mammalian cells and tissues [ 15 – 18 ]. Yuan et al [ 7 ] actually showed that supplementing DMSO formulations with ficoll allowed preservation of pluripotent stem cells for up to one year at −80°C, whereas formulations without ficoll were not effective.…”

Comprehensive evaluation of different cryopreservation methods used for the successful storage of human Wharton's Jelly tissue

“…The cryopreservation of MSCs, a widely applied method, could be used as an alternative approach in order to preserve the cells intact for a long-time period at -196 o C [10]. Ideally, the MSCs could be recovered at any chosen time point and can be used in translation medicine [10].…”

“…Ideally, the MSCs could be recovered at any chosen time point and can be used in translation medicine [10]. Although, when these cells are recovered, it is necessary to be expanded in vitro, thus prolonging their culture even more.…”