Thawed cryopreserved synovial mesenchymal stem cells show comparable effects to cultured cells in the inhibition of osteoarthritis progression in rats

Kiyotaka Horiuchi,Kunikazu Tsuji,Hideyuki Koga,Kentaro Endo,Nobutake Ozeki,Ichiro Sekiya,Masako Akiyama,Hisako Katano,Mitsuru Mizuno

doi:10.1038/s41598-021-89239-8

Kiyotaka Horiuchi, Kunikazu Tsuji + Show 7 more

Open Access

https://doi.org/10.1038/s41598-021-89239-8

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Journal: Scientific Reports	Publication Date: May 6, 2021
Citations: 7	License type: open-access

Affiliation: Tokyo Medical and Dental University

Abstract

Intra-articular injections of mesenchymal stem cells (MSCs) can inhibit the progression of osteoarthritis (OA). 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. Cultured rat synovial MSCs or thawed MSCs were compared for in vitro viability and properties. The inhibitory effect of thawed MSCs on OA progression was evaluated by injecting cryopreservation fluid and thawed MSCs in meniscectomized rats. Cartilage degeneration was assessed using gross finding and histological scores. Cultured MSCs were then injected into one knee and thawed MSCs into the contralateral knee of the same individual to compare their effects. Cultured MSCs and MSCs thawed after cryopreservation had comparable in vitro colony formation and chondrogenic potentials. In the rat OA model, the gross finding and histological scores were significantly lower in the thawed MSC group than in the cryopreservation fluid group at 8 weeks. Finally, cartilage degeneration did not differ significantly after injection of cultured and thawed MSCs. In conclusion, thawed MSCs showed comparable inhibitory effects on OA progression to cultured MSCs.

Highlights

Intra-articular injections of mesenchymal stem cells (MSCs) can inhibit the progression of osteoarthritis (OA)
MSCs thawed after cryopreservation in 95% fetal bovine serum (FBS) with 5% dimethyl sulfoxide (DMSO) or 100% FBS were compared with MSCs without cryopreservation (Fig. 1A)
In vitro viability was not significantly different in MSCs thawed after cryopreservation in 95% FBS with 5% DMSO versus cultured MSCs (Fig. 1B), but cellular dehydrogenase activity and lactate dehydrogenase activity were significantly lower in the thawed MSCs as well as in MSCs thawed after cryopreservation in 100% FBS (Fig. 1C,D)

Summary

Introduction

Intra-articular injections of mesenchymal stem cells (MSCs) can inhibit the progression of osteoarthritis (OA). The inhibitory effect of thawed MSCs on OA progression was evaluated by injecting cryopreservation fluid and thawed MSCs in meniscectomized rats. Cultured MSCs and MSCs thawed after cryopreservation had comparable in vitro colony formation and chondrogenic potentials. Cartilage degeneration did not differ significantly after injection of cultured and thawed MSCs. In conclusion, thawed MSCs showed comparable inhibitory effects on OA progression to cultured MSCs. Osteoarthritis (OA) is the most common joint disease in the w orld[1] and a leading cause of disability worldwide. A growing number of papers have compared cultured and thawed MSCs for their properties as MSCs and their therapeutic effects on some diseases. ND, not detected; NS not significant; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by repeated measures one-way ANOVA followed by Tukey’s multiple comparisons (B–D,K), Kruskal–Wallis test followed by Dunn’s multiple comparisons (F–H) or Student’s t-test between two unpaired groups (J)

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