Mizuno Mitsuru, Katano Hisako, Otabe Koji, Komori Keiichiro, Kohno Yuji, Fujii Shizuka, Ozeki Nobutake, Horie Masafumi, Tsuji Kunikazu, Koga Hideyuki, Muneta Takeshi, Sekiya Ichiro
Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.
Department of Joint Surgery and Sports Medicine, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Stem Cell Res Ther. 2017 Jun 13;8(1):144. doi: 10.1186/s13287-017-0596-0.
In 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.
Human 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.
After 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.
The viability and chondrogenic potential of synovial MSCs were maintained when the cells were suspended in human serum at 4 and 13 °C.
在我们的临床实践中，我们进行自体滑膜间充质干细胞（MSCs）移植用于软骨和半月板再生医学。确保临床疗效的最重要问题之一涉及滑膜间充质干细胞从处理设施运输到诊所的过程。完全人血清（100%人血清）是一种有吸引力的候选材料，可用于悬浮滑膜间充质干细胞以便在运输过程中保存。本研究的目的是调查完全人血清是否能维持间充质干细胞的活力和软骨形成潜力，并研究保存人滑膜间充质干细胞的最佳温度条件。
在全膝关节置换术中从14例骨关节炎供体的膝关节采集人滑膜。第2代滑膜间充质干细胞以200万个细胞/100μL的浓度悬浮于林格氏液或完全人血清中，分别在4℃、13℃和37℃保存48小时。在保存前后对这些细胞进行活细胞率、细胞表面标志物表达、代谢活性、增殖以及成脂、钙化和软骨形成分化潜能分析。
保存后，在4℃和13℃时，滑膜间充质干细胞在人血清中的活细胞率高于在林格氏液中的活细胞率。在4℃和13℃保存在人血清中的滑膜间充质干细胞也保持了较高比例的碘化丙啶和膜联蛋白V细胞。在4℃和13℃保存的细胞中，间充质干细胞表面标志物表达未改变。在4℃和13℃保存在人血清中的细胞代谢活性得以维持，而在其他条件下显著降低。当在4℃和13℃保存在人血清中时，重新接种的间充质干细胞保留了其增殖能力。在每种保存条件下的细胞中均可观察到成脂和钙化潜力，而仅在4℃和13℃保存在人血清中的细胞中保留了软骨形成潜力。
当滑膜间充质干细胞悬浮于4℃和13℃的人血清中时，其活力和软骨形成潜力得以维持。
