Abstract
BackgroundSynovial mesenchymal stem cells (MSCs) are an attractive cell source for cartilage and meniscus regeneration. The optimum cryopreservation medium has not been determined, but dimethylsulfoxide (DMSO) should be excluded, if possible, because of its toxicity. The purposes of our study were to examine the possible benefits of higher concentrations of serum and the effectiveness of 100% serum (without DMSO) for the cryopreservation of synovial MSCs.MethodsHuman synovium was harvested from the knees of four donors with osteoarthritis during total knee arthroplasty. Synovial MSCs (8 × 105 cells) were suspended in 400 μL medium and used as a Time 0 control. The same number of synovial MSCs was also suspended in 400 μL α-MEM medium containing 10% fetal bovine serum (FBS) (5% DMSO, and 1% antibiotic), 95% FBS (and 5% DMSO), or 100% FBS (no DMSO) and cryopreserved at − 80 °C for 7 days. After thawing, the cell suspensions (1.5 μL; 3 × 103 cells) were cultured in 60 cm2 dishes for 14 days for colony formation assays. Additional 62.5 μL samples of cell suspensions (1.25 × 105 cells) were added to tubes and cultured for 21 days for chondrogenesis assays.ResultsColony numbers were significantly higher in the Time 0 and 95% FBS groups than in the 10% FBS group (n = 24). Colony numbers were much lower in the 100% FBS group than in the other three groups. The cell numbers per dish reflected the colony numbers. Cartilage pellet weights were significantly heavier in the 95% FBS group than in the 10% FBS group, whereas no difference was observed between the Time 0 and the 95% FBS groups (n = 24). No cartilage pellets formed at all in the 100% FBS group.ConclusionSynovial MSCs cryopreserved in 95% FBS with 5% DMSO maintained their colony formation and chondrogenic abilities to the same levels as observed in the cells before cryopreservation. Synovial MSCs cryopreserved in 100% FBS lost their colony formation and chondrogenic abilities.
Highlights
Synovial mesenchymal stem cells (MSCs) are an attractive cell source for cartilage and meniscus regeneration
We recently reported that synovial MSCs maintained their viability as well as their chondrogenic ability for 48 h when preserved in 100% human serum at 4 °C or 13 °C [7]
We examined the effect of the cryopreservation medium composition on the maintenance of the colony formation and chondrogenic abilities of synovial MSCs
Summary
Synovial mesenchymal stem cells (MSCs) are an attractive cell source for cartilage and meniscus regeneration. The purposes of our study were to examine the possible benefits of higher concentrations of serum and the effectiveness of 100% serum (without DMSO) for the cryopreservation of synovial MSCs. Human synovial mesenchymal stem cells (MSCs) are a promising cell source for regeneration of cartilage and meniscus [1]. We recently reported that synovial MSCs maintained their viability as well as their chondrogenic ability for 48 h when preserved in 100% human serum at 4 °C or 13 °C [7] This suggests that higher concentrations of serum in the cryopreservation medium improves cell viability
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