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.
Highlights
In our clinical practice, we perform transplantations of autologous synovial mesenchymal stem cells (MSCs) for cartilage and meniscus regenerative medicine
Apoptosis of synovial MSCs 48 hours after preservation According to forward scatter (FSC) analyses, synovial MSCs appeared to shift to a smaller size profile 48 h after preservation, cells preserved at 37 °C (Fig. 4a and b)
We investigated whether complete human serum maintained the viability and chondrogenic potential, and examined the ideal temperature for the 48-h preservation, of human synovial MSCs
Summary
We perform transplantations of autologous synovial mesenchymal stem cells (MSCs) for cartilage and meniscus regenerative medicine. 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. It has been shown that mesenchymal stem cells (MSCs) derived from synovium readily expand in vitro in human serum and differentiate into cartilage and meniscus tissue when transplanted into these defects in vivo [1, 2]. Synovial tissue was harvested arthroscopically and enzymatically digested, synovial cells were cultured with 10% autologous human serum for 14 days, and synovial MSCs were suspended in 0.5 mL glucose acetate Ringer’s solution and transplanted arthroscopically onto the cartilage and/or meniscus defects (Fig. 1a). We wanted to determine the best suspension conditions, temperature, and medium for the transport of synovial MSCs to guarantee patient safety
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