Complete human serum maintains viability and chondrogenic potential of human synovial stem cells: suitable conditions for transplantation

Abstract

BackgroundIn our clinical practice, we perform transplantations of autologous synovial mesenchymal stem cells (MSCs) for cartilage and meniscus regenerative medicine. One of the most important issues to ensuring clinical efficacy involves the transport of synovial MSCs from the processing facility to the clinic. Complete human serum (100% human serum) is an attractive candidate material in which to suspend synovial MSCs for their preservation during transport. The purpose of this study was to investigate whether complete human serum maintained MSC viability and chondrogenic potential and to examine the optimal temperature conditions for the preservation of human synovial MSCs.MethodsHuman synovium was harvested from the knees of 14 donors with osteoarthritis during total knee arthroplasty. Passage 2 synovial MSCs were suspended at 2 million cells/100 μL in Ringer’s solution or complete human serum at 4, 13, and 37 °C for 48 h. These cells were analyzed for live cell rates, cell surface marker expression, metabolic activity, proliferation, and adipogenic, calcification, and chondrogenic differentiation potentials before and after preservation.ResultsAfter preservation, synovial MSCs maintained higher live cell rates in human serum than in Ringer’s solution at 4 and 13 °C. Synovial MSCs preserved in human serum at 4 and 13 °C also maintained high ratios of propidium iodide– and annexin V– cells. MSC surface marker expression was not altered in cells preserved at 4 and 13 °C. The metabolic activities of cells preserved in human serum at 4 and 13 °C was maintained, while significantly reduced in other conditions. Replated MSCs retained their proliferation ability when preserved in human serum at 4 and 13 °C. Adipogenesis and calcification potential could be observed in cells preserved in each condition, whereas chondrogenic potential was retained only in cells preserved in human serum at 4 and 13 °C.ConclusionThe viability and chondrogenic potential of synovial MSCs were maintained when the cells were suspended in human serum at 4 and 13 °C.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0596-0) contains supplementary material, which is available to authorized users.

Cryopreservation in 95% serum with 5% DMSO maintains colony formation and chondrogenic abilities in human synovial mesenchymal stem cells

“…In the present study, the culture medium for the 10% FBS group contained 1% antibiotics, whereas the media for the 95 and 100% FBS groups did not. We initially hypothesized that 100% FBS would function as a cryopreservation medium because 100% FBS worked as a preservation medium [7]. We also wanted to examine the additional effect of DMSO (i.e., in the 95% FBS group).…”

“…Our colony formation and chondrogenesis assays were conducted by adding cell suspensions directly to dishes or tubes, without first determining the numbers of viable cells, as we considered this a more accurate method for evaluating cell viability. Our previous studies indicated that the statistical dispersions of the live cell rate, as determined by live/dead assays, and the apoptosis rate, as determined by flow cytometry, were relatively wide for human synovial MSCs after 48 h of preservation in Ringer’s solution or in complete human serum at 4, 13, and 37 °C [7].…”

“…However, an optimum cryopreservation medium has yet to be identified. 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.…”

Cardiomyocyte differentiation of mesenchymal stem cells from bone marrow: new regulators and its implications

“…As the expansion of MSCs performed in cell culture medium with FBS has several problems including viral, bacterial, and prion [ 80 ], autologous serum has been considered. Autologous serum could regulate the proliferation and differentiation of MSCs [ 81 ] and enhance the cell viability. Chachques et al [ 82 ] had transplanted autologous myoblasts which were cultured in autologous serum medium into LV infarcted patients to repair the jeopardized myocardium.…”

Combating Osteoarthritis through Stem Cell Therapies by Rejuvenating Cartilage: A Review

“…SMSCs are also isolated from hip joints; however, those isolated from knee joints have shown a better chondrogenic potential [ 95 ]. These MSCs can also be preserved in complete human serum at 4 or 13°C without significantly affecting their viability and chondrogenic potential [ 96 ]. In an interesting research, the exosomes derived from SMSC-140s promoted chondrogenesis without affecting the quality of ECM [ 97 ].…”