Preservation media, durations and cell concentrations of short-term storage affect key features of human adipose-derived mesenchymal stem cells for therapeutic application

Abstract

BackgroundAdipose-derived mesenchymal stem cells (ADSCs) have shown great potential in the treatment of various diseases. However, the optimum short-term storage condition of ADSCs in 2∼8 °C is rarely reported. This study aimed at optimizing a short-term storage condition to ensure the viability and function of ADSCs before transplantation.MethodsPreservation media and durations of storage were evaluated by cell viability, apoptosis, adhesion ability and colony-forming unit (CFU) capacity of ADSCs. The abilities of cell proliferation and differentiation were used to optimize cell concentrations. Optimized preservation condition was evaluated by cell surface markers, cell cycle and immunosuppressive capacity.ResultsA total of 5% human serum albumin in multiple electrolytes (ME + HSA) was the optimized medium with high cell viability, low cluster rate, good adhesion ability and high CFU capacity of ADSCs. Duration of storage should be limited to 24 h to ensure the quality of ADSCs before transplantation. A concentration of 5 × 106 cells/ml was the most suitable cell concentration with low late stage apoptosis, rapid proliferation and good osteogenic and adipogenic differentiation ability. This selected condition did not change surface markers, cell cycle, indoleamine 2, 3-dioxygenase 1 (IDO1) gene expression and kynurenine (Kyn) concentration significantly.DiscussionIn this study, ME + HSA was found to be the best medium, most likely due to the supplement of HSA which could protect cells, the physiological pH (7.4) of ME and sodium gluconate ingredient in ME which could provide energy for cells. Duration should be limited to 24 h because of reduced nutrient supply and increased waste and lactic acid accumulation during prolonged storage. To keep cell proliferation and limit lactic acid accumulation, the proper cell concentration is 5× 106 cells/ml. Surface markers, cell cycle and immunosuppressive capacity did not change significantly after storage using the optimized condition, which confirmed our results that this optimized short-term storage condition of MSCs has a great potential for the application of cell therapy.

Modelling of the SDF-1/CXCR4 regulated in vivo homing of therapeutic mesenchymal stem/stromal cells in mice

“…Mesenchymal stem/stromal cells (MSCs) are excellent candidates for use in tissue repair and regeneration (Fu et al, 2018; Niclis et al, 2017; Squillaro, Peluso & Galderisi, 2016; Zhang et al, 2017). Human MSCs can be harvested from a range of tissues (bone marrow and adipose are common sources) with few ethical issues; and these cells can be expanded in number for use on clinical scales within a short time period (Parekkadan & Milwid, 2010; Rohart et al, 2016; Zhang et al, 2017). Similar to the use of pharmacokinetics for drug development, the aim of elucidating in vivo kinetics of MSCs is to predict and enhance their therapeutic potential, as well as to minimize adverse effects.…”

Effects of storage culture media, temperature and duration on human adipose‑derived stem cell viability for clinical use

“…In addition to the temperature, the preservation medium is also of critical significance. Several reports have concluded that an environment that mimics that of the native cell, containing 0.9% NaCl, 5% glucose and albumin or human serum (HS), is the most appropriate for cell survival (14). Other preservation media have also been studied, such as DMSO; however, the clinical use of DMSO causes diverse problems, including leukoencephalopathy, nausea, vomiting and potential renal function decrease (15).…”

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“…There is a growing interest in MSCs in the context of regenerative medicine for treating injured organs (Jiang et al, 2018;Squillaro et al, 2016;Zhang et al, 2017). Therefore, understanding the kinetics, including the homing of MSCs, is becoming crucial to improve treatment outcomes.…”

“…Mesenchymal stem/stromal cells (MSCs) are excellent candidates for use in tissue repair and regeneration (Jiang et al, 2018;Niclis et al, 2017;Squillaro et al, 2016;Zhang et al, 2017).…”

“…Human MSCs can be harvested from a range of tissues (bone marrow and adipose are common sources) with few ethical issues; and MSC-based therapies have few adverse events (Parekkadan et al, 2010;Rohart et al, 2016;Zhang et al, 2017). Similar to the use of pharmacokinetics for drug development, the aim of elucidating in vivo kinetics of MSCs is to predict and enhance their therapeutic potential.…”