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Open Access 01-12-2019 | Research article

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

Authors: Ryota Fujisawa, Mitsuru Mizuno, Hisako Katano, Koji Otabe, Nobutake Ozeki, Kunikazu Tsuji, Hideyuki Koga, Ichiro Sekiya

Published in: BMC Musculoskeletal Disorders | Issue 1/2019

Abstract

Background

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.

Methods

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.

Results

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.

Conclusion

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.

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Title: Cryopreservation in 95% serum with 5% DMSO maintains colony formation and chondrogenic abilities in human synovial mesenchymal stem cells
Authors: Ryota Fujisawa
Mitsuru Mizuno
Hisako Katano
Koji Otabe
Nobutake Ozeki
Kunikazu Tsuji
Hideyuki Koga
Ichiro Sekiya
Publication date: 01-12-2019
Publisher: BioMed Central
Published in: BMC Musculoskeletal Disorders / Issue 1/2019
Electronic ISSN: 1471-2474
DOI: https://doi.org/10.1186/s12891-019-2700-3

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

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

Abstract

Background

Methods

Results

Conclusion

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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