Cryopreservation in 95% serum with 5% DMSO maintains colony formation and chondrogenic abilities in human synovial mesenchymal stem cells
Open Access
- 6 July 2019
- journal article
- research article
- Published by Springer Science and Business Media LLC in BMC Musculoskeletal Disorders
- Vol. 20 (1), 1-9
- https://doi.org/10.1186/s12891-019-2700-3
Abstract
Synovial 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. Human 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. Colony 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. Synovial 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.Keywords
Funding Information
- Japan Society for the Promotion of Science (16H06262)
- Japan Agency for Medical Research and Development (JP16bm0504001)
This publication has 44 references indexed in Scilit:
- Hydroxyethylstarch in cryopreservation – Mechanisms, benefits and problemsTransfusion and Apheresis Science, 2012
- Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem CellsStem Cells and Development, 2010
- Increased proliferation of human synovial mesenchymal stem cells with autologous human serum: Comparisons with bone marrow mesenchymal stem cells and with fetal bovine serumArthritis & Rheumatism, 2008
- Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statementCytotherapy, 2006
- Cryopreservation of Stem Cells Using Trehalose: Evaluation of the Method Using a Human Hematopoietic Cell LineStem Cells and Development, 2004
- TOXICOLOGIC UPDATE OF DIMETHYL SULFOXIDEAnnals of the New York Academy of Sciences, 1983
- Cryopreservation ofDigitalis lanataCell CulturesPlanta Medica, 1982
- Cryopreservation of platelet concentrates using glycerol-glucoseTransfusion, 1982
- Cryopreservation of human platelets with polyvinylpyrrolidoneTransfusion, 1981
- SAFETY OF DIMETHYLSULPHOXIDEThe Lancet, 1981