Efficient cryopreservation of human mesenchymal stem cells using silkworm hemolymph-derived proteins

Kim, Sunmi; Yun, Chang-Koo; Park, Jinho; Hwang, Jung Wook; Kim, Z-Hun; Choi, Yong‐Soo

doi:10.1002/term.2116

Cited by 3 publications

(3 citation statements)

References 37 publications

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“…Had we conducted the cryopreservation in a liquid nitrogen tank for longer durations, our outcomes might have been different. Kim et al reported that the viability of human umbilical cord MSCs cryopreserved in a liquid nitrogen tank for 1 year was comparable to that of cells cryopreserved for 7 days [37]. Abbruzzese at al.…”

Section: Discussionmentioning

confidence: 99%

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

Fujisawa

Mizuno

Katano

et al. 2019

BMC Musculoskelet Disord

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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 × 10 5 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 × 10 3 cells) were cultured in 60 cm 2 dishes for 14 days for colony formation assays. Additional 62.5 μL samples of cell suspensions (1.25 × 10 5 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. Electronic supplementary material The online version of this article (10.1186/s12891-019-2700-3) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

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

Fujisawa

Mizuno

Katano

et al. 2019

BMC Musculoskelet Disord

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“…A longer period of cryopreservation could lead to different results. However, the viability of human umbilical cord MSCs cryopreserved for 1 year was comparable to that of cells cryopreserved for 7 days [31], suggesting that the results would not change even if the synovial MSCs were cryopreserved for 1 year.…”

Section: Discussionmentioning

confidence: 94%

Thawed Cryopreserved Synovial Mesenchymal Stem Cells Show Comparable Effects to Cultured Cells in the Inhibition of Osteoarthritis Progression in Rats

Horiuchi¹,

Ozeki²,

Endo³

et al. 2020

Preprint

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Background Intra-articular injections of mesenchymal stem cells (MSCs) can inhibit the progression of osteoarthritis (OA) in animal studies. Previous reports have used cultured MSCs, but the ability to use thawed cryopreserved MSC stocks would be highly advantageous. Our purpose was to elucidate whether thawed cryopreserved MSCs show comparable inhibitory effects on OA progression in rats to those obtained with cultured MSCs. Methods Cultured rat synovial MSCs or thawed MSCs were compared for in vitro viability and properties. The activity of synovial MSCs derived from rats expressing luciferase was compared by the luminescence intensity both in vitro and in vivo. The inhibitory effect of cultured MSCs on OA progression was evaluated by injecting PBS and cultured MSCs into the right and left knees, respectively, of the same individual meniscectomized rats. A similar test evaluated cryopreservation fluid (95% FBS with 5% DMSO; termed the 95% FBS group) versus thawed MSCs. Cartilage degeneration was assessed at 4 and 8 weeks using gross finding and histological scores. The results were used to set the primary outcome and sample size. Cultured MSCs were then injected into one knee and thawed MSCs into the contralateral knee of the same individual to compare their effects.Results Cultured MSCs and MSCs thawed after cryopreservation in 95% FBS with 5% DMSO had comparable in vitro viability, colony formation, and chondrogenic potential. The luminescence intensity was comparable between the two MSC preparations. In the rat OA model, the gross findings and histological scores for tibial cartilage did not differ at four weeks but were significantly lower in the cultured MSC group than in the PBS group and significantly lower in the thawed MSC group than in the 95% FBS group at eight weeks. These results established the tibial cartilage histological score at eight weeks as the primary outcome and a sample size of nine for comparing cultured versus thawed MSCs. The tibial cartilage histological scores did not differ significantly at eight weeks after injection of cultured and thawed MSCs into the opposite knees of the same individuals.ConclusionsThawed MSCs showed comparable inhibitory effects on OA progression to cultured MSCs.

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“…1 a Schematic diagram for two types of cryoinjuries occurring during cryopreservation of cells; b schematic diagram for various biotechnology techniques used in cryopreservation and their method of protection extracellular cryopreservation. However, FBS is derived from animals and has risk of inducing viral infections and immunogenicity inpatients [20,21].…”

Section: Introductionmentioning

confidence: 99%

Advanced Biotechnology for Cell Cryopreservation

Yang

Gao

Liu

et al. 2019

Trans. Tianjin Univ.

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Cell cryopreservation has evolved as an important technology required for supporting various cell-based applications, such as stem cell therapy, tissue engineering, and assisted reproduction. Recent times have witnessed an increase in the clinical demand of these applications, requiring urgent improvements in cell cryopreservation. However, cryopreservation technology suffers from the issues of low cryopreservation efficiency and cryoprotectant (CPA) toxicity. Application of advanced biotechnology tools can significantly improve post-thaw cell survival and reduce or even eliminate the use of organic solvent CPAs, thus promoting the development of cryopreservation. Herein, based on the different cryopreservation mechanisms available, we provide an overview of the applications and achievements of various biotechnology tools used in cell cryopreservation, including trehalose delivery, hydrogel-based cell encapsulation technique, droplet-based cell printing, and nanowarming, and also discuss the associated challenges and perspectives for future development.

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Efficient cryopreservation of human mesenchymal stem cells using silkworm hemolymph-derived proteins

Cited by 3 publications

References 37 publications

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

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

Thawed Cryopreserved Synovial Mesenchymal Stem Cells Show Comparable Effects to Cultured Cells in the Inhibition of Osteoarthritis Progression in Rats

Advanced Biotechnology for Cell Cryopreservation

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