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Article ; Online: Characteristics of MSCs in Synovial Fluid and Mode of Action of Intra-Articular Injections of Synovial MSCs in Knee Osteoarthritis

Ichiro Sekiya / Hisako Katano / Nobutake Ozeki

International Journal of Molecular Sciences, Vol 22, Iss 2838, p

2021 Volume 2838

Abstract: We have been studying mesenchymal stem cells (MSCs) in synovial fluid and the intra-articular injection of synovial MSCs in osteoarthritis (OA) knees. Here, mainly based on our own findings, we overview the characteristics of endogenous MSCs in the ... ...

Abstract	We have been studying mesenchymal stem cells (MSCs) in synovial fluid and the intra-articular injection of synovial MSCs in osteoarthritis (OA) knees. Here, mainly based on our own findings, we overview the characteristics of endogenous MSCs in the synovial fluid of OA knees and their mode of action when injected exogenously into OA knees. Many MSCs similar to synovial MSCs were detected in the synovial fluid of human OA knees, and their number correlated with the radiological OA grade. Our suspended synovium culture model demonstrated the release of MSCs from the synovium through a medium into a non-contacting culture dish. In OA knees, endogenous MSCs possibly mobilize in a similar manner from the synovium through the synovial fluid and act protectively. However, the number of mobilized MSCs is limited; therefore, OA progresses in its natural course. Synovial MSC injections inhibited the progression of cartilage degeneration in a rat OA model. Injected synovial MSCs migrated into the synovium, maintained their MSC properties, and increased the gene expressions of TSG-6, PRG-4, and BMP-2. Exogenous synovial MSCs can promote anti-inflammation, lubrication, and cartilage matrix synthesis in OA knees. Based on our findings, we have initiated a human clinical study of synovial MSC injections in OA knees.
Keywords	MSC ; synovial fluid ; mode of action ; injection ; synovium ; knee ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	616
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Stress and motivation of cell processing operators

Mitsuru Mizuno / Yoshimi Sugahara / Daisuke Iwayama / Noe Miyashita / Hisako Katano / Ichiro Sekiya

Regenerative Therapy, Vol 21, Iss , Pp 547-

A pilot study of an online questionnaire survey

2022 Volume 552

Abstract: Introduction: Cell processing operators (CPOs) are one of the most important human assets for the industrial development of regenerative therapy since cell processing is still carried out manually. However, investigating the psychological aspects of CPOs ...

Abstract	Introduction: Cell processing operators (CPOs) are one of the most important human assets for the industrial development of regenerative therapy since cell processing is still carried out manually. However, investigating the psychological aspects of CPOs to take advantage of this important human asset has not been done. Thus, the objectives of the current study were, first, to investigate the attributes of CPOs, second, to identify the factors that give CPOs mental stress, and third, to identify the motivational factors of CPOs. Methods: An online questionnaire was conducted to investigate the attributes of participants, stresses, and motivations; the questionnaire included multiple-choice questions. A survey questionnaire was distributed to 64 operators in 31 CPFs, including universities, clinics, and pharmaceutical companies, from December 2021 to February 2022. Participants responded to the online survey via Microsoft Forms. Results: Approximately half of the CPOs had been employed for two years or less, and they were more likely to experience various stressful situations than CPOs with three or more years of experience. In particular, the wear of dust-free clothing and work in a unique environment of a clean room, where it is difficult to take breaks due to the nature of the closed space, were considered to be particularly stressful situations. However, these CPOs have high motivations based on their pride in being involved in cutting-edge medical care, which is highly expected by society, and in contributing to patient care. Conclusions: While stress with training and education programs for CPOs is an issue that can be resolved through in-house training, other issues that are difficult to resolve immediately, such as mechanization to relieve the stress of human manual operations, were identified. Continued efforts to analyze and alleviate the stress factors identified in this study are expected to improve the quality of CPOs’ work and maintain the important human asset of CPOs.
Keywords	Cell processing operators (CPOs) ; Online questionnaire survey ; Stress ; Motivation ; Cell processing facilities (CPFs) ; Medicine (General) ; R5-920 ; Cytology ; QH573-671
Subject code	150
Language	English
Publishing date	2022-12-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Three-dimensional MRI shows cartilage defect extension with no separation from the meniscus in women in their 70 s with knee osteoarthritis

Hisako Katano / Nobutake Ozeki / Hideyuki Koga / Makoto Tomita / Kenji Suzuki / Jun Masumoto / Ichiro Sekiya

Scientific Reports, Vol 12, Iss 1, Pp 1-

2022 Volume 10

Abstract: Abstract The positional relationship between cartilage defects and the meniscus is poorly understood for osteoarthritis of the knee. Our purpose was to clarify how cartilage defects extend and their association with the meniscus location during ... ...

Abstract	Abstract The positional relationship between cartilage defects and the meniscus is poorly understood for osteoarthritis of the knee. Our purpose was to clarify how cartilage defects extend and their association with the meniscus location during osteoarthritis progression. The subjects were women in their 70 s who were registered in the Kanagawa Knee Study. We obtained 3D MRI images of the tibial surfaces with menisci in subjects with cartilage area ratios < 0.95 and examined the morphological association between cartilage defects and the medial meniscus (MM) by viewing the defects according to the cartilage area ratio at the medial tibial region. Of the 561 Kanagawa Knee Study subjects, 45 were included in the analyses, and 11 had a cartilage area ratio < 0.95 at the medial tibia. Significant differences were observed in the localization of cartilage defects among 9 subregions, with cartilage defects occurring predominantly in the middle external subregion. The inner margin of the MM contacted the cartilage defect in 7 knees and crossed the cartilage defect in 4 knees but was never found separated from the cartilage defect. The cartilage defects occurred from the middle external subregion and extended to the surrounding area without separating from the inner margin of the MM. Trial registration UMIN, UMIN000032826; 1 September 2018.
Keywords	Medicine ; R ; Science ; Q
Subject code	616
Language	English
Publishing date	2022-03-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Alterations in cartilage quantification before and after injections of mesenchymal stem cells into osteoarthritic knees

Ichiro Sekiya / Hisako Katano / Mitsuru Mizuno / Hideyuki Koga / Jun Masumoto / Makoto Tomita / Nobutake Ozeki

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 14

Abstract: Abstract Several studies have reported improvement in knee pain following mesenchymal stem cell (MSC) injections for knee osteoarthritis (OA). We developed a novel 3D magnetic resonance imaging (MRI) analysis software program that provides “projected ... ...

Abstract	Abstract Several studies have reported improvement in knee pain following mesenchymal stem cell (MSC) injections for knee osteoarthritis (OA). We developed a novel 3D magnetic resonance imaging (MRI) analysis software program that provides “projected cartilage area ratios” for automatic detection of changes in cartilage amounts. The primary objective of this prospective interventional study was to compare alterations in the projected cartilage area ratio (thickness ≥ 1.5 mm) at the femoral posteromedial region between 30 weeks before and 30 weeks after synovial MSC injections. Secondary objectives were to assess the clinical scores and safety of MSC injections. Patients with OA who complained of knee pain underwent autologous synovial MSC injections into the knee at time 0 and again 15 weeks later. MRI examinations were performed at − 30, − 15, − 1, and 30 weeks. Patients showing < 3% decreases in the projected cartilage area ratio (thickness ≥ 1.5 mm) at the femoral the posteromedial region from − 30 weeks to − 15 weeks were excluded from the study. The Lysholm Knee Score, Knee Injury and Osteoarthritis Outcome Scale (KOOS), and Numerical Rating Scale (NRS) scores were evaluated at − 30, − 15, − 5, − 2, 0, 5, 10, 15, 20, 25, and 30 weeks. Five patients were excluded because 3D MRI analysis showed no cartilage loss at − 15 weeks. Ultimately, eight OA patients underwent MSC injections. The projected cartilage area ratio significantly decreased by 0.07 in the 30 weeks before MSC injections (p = 0.01), but no further decreases occurred in the 30 weeks after MSC injections. The projected cartilage area ratio at the femoral posteromedial region showed a significant difference between 30 weeks before and 30 weeks after MSC injections. The Lysholm Knee Score, KOOS, and NRS values improved significantly after the injections. MSC injection could not be ruled out as the cause of two adverse events: transient knee pain and itching in both hands. Fully automatic 3D MRI analysis showed that synovial MSC injections ...
Keywords	Medicine ; R ; Science ; Q
Subject code	796
Language	English
Publishing date	2021-07-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs

Mitsuru Mizuno / Takahisa Matsuzaki / Nobutake Ozeki / Hisako Katano / Hideyuki Koga / Takanori Takebe / Hiroshi Y. Yoshikawa / Ichiro Sekiya

Stem Cell Research & Therapy, Vol 13, Iss 1, Pp 1-

2022 Volume 16

Abstract: Abstract Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the ... ...

Abstract	Abstract Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed to identify the biological and physical factors involved in the differences in freeze–thaw tolerance between MSCs and HUVECs. Materials and methods For biological analysis, MSC and HUVEC viability after freeze-thawing and alteration of gene expression in response to dimethyl sulfoxide (DMSO, a cryoprotectant) were quantitatively evaluated. For physical analysis, the cell membrane fluidity of MSCs and HUVECs before and after DMSO addition was assessed using a histogram for generalized polarization frequency. Results HUVECs showed lower live cell rates and higher gene expression alteration related to extracellular vesicles in response to DMSO than MSCs. Fluidity measurements revealed that the HUVEC membrane was highly fluidic and sensitive to DMSO compared to that of MSCs. Addition of CAY10566, an inhibitor of stearoyl-coA desaturase (SCD1) that produces highly fluidic desaturated fatty acids, decreased the fluidity of HUVECs and increased their tolerance to DMSO. The combination of CAY10566 and antioxidant glutathione (GSH) treatment improved HUVEC viability from 57 to 69%. Membrane fluidity alteration may thus contribute to pore-induced DMSO influx into the cytoplasm and reactive oxygen species production, leading to greater cytotoxicity in HUVECs, which have low antioxidant capacity. Conclusions Differences in freeze–thaw tolerance originate from differences in the cell membranes with respect to fluidity and antioxidant capacity. These findings provide a basis for analyzing cell biology and membrane-physics to establish appropriate long-term preservation methods aimed at promoting transplantation therapies.
Keywords	Cell membrane fluidity ; ROS resistance ; Cryopreserve ; Mesenchymal stem cells ; Human umbilical vein endothelial cells ; Medicine (General) ; R5-920 ; Biochemistry ; QD415-436
Subject code	571
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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

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

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 14

Abstract: 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 ... ...

Abstract	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.
Keywords	Medicine ; R ; Science ; Q
Subject code	610 ; 571
Language	English
Publishing date	2021-05-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Optimal Pore Size of Honeycomb Polylactic Acid Films for In Vitro Cartilage Formation by Synovial Mesenchymal Stem Cells

Misaki Yagi / Mitsuru Mizuno / Ryota Fujisawa / Hisako Katano / Kentaro Endo / Nobutake Ozeki / Yuriko Sakamaki / Hideyuki Koga / Ichiro Sekiya

Stem Cells International, Vol

2021 Volume 2021

Abstract: Background. Tissue engineering of cartilage requires the selection of an appropriate artificial scaffold. Polylactic acid (PLA) honeycomb films are expected to be highly biodegradable and cell adhesive due to their high porosity. The purpose of this ... ...

Abstract	Background. Tissue engineering of cartilage requires the selection of an appropriate artificial scaffold. Polylactic acid (PLA) honeycomb films are expected to be highly biodegradable and cell adhesive due to their high porosity. The purpose of this study was to determine the optimal pore size of honeycomb PLA films for in vitro cartilage formation using synovial mesenchymal stem cells (MSCs). Methods. Suspensions of human synovial MSCs were plated on PLA films with different pore sizes (no pores, or with 5 μm or 20 μm pores) and then observed by scanning electron microscopy. The numbers of cells remaining in the film and passing through the film were quantified. One day after plating, the medium was switched to chondrogenic induction medium, and the films were time-lapse imaged and observed histologically. Results. The 5 μm pore film showed MSCs with pseudopodia that extended between several pores, while the 20 μm pore film showed MSC bodies submerged into the pores. The number of adhered MSCs was significantly lower for the film without pores, while the number of MSCs that passed through the film was significantly higher for the 20 μm pore film. MSCs that were induced to form cartilage peeled off as a sheet from the poreless film after one day. MSCs formed thicker cartilage at two weeks when growing on the 5 μm pore films than on the 20 μm pore films. Conclusions. Honeycomb PLA films with 5 μm pores were suitable for in vitro cartilage formation by synovial MSCs.
Keywords	Internal medicine ; RC31-1245
Subject code	620
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Hindawi Limited
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Time-lapse image analysis for whole colony growth curves and daily distribution of the cell number per colony during the expansion of mesenchymal stem cells

Mitsuru Mizuno / Hisako Katano / Yuri Shimozaki / Sho Sanami / Nobutake Ozeki / Hideyuki Koga / Ichiro Sekiya

Scientific Reports, Vol 9, Iss 1, Pp 1-

2019 Volume 9

Abstract: Abstract Mesenchymal stem cells from the synovium (synovial MSCs) are attractive for cartilage and meniscus regeneration therapy. We developed a software program that can distinguish individual colonies and automatically count the cell number per colony ... ...

Abstract	Abstract Mesenchymal stem cells from the synovium (synovial MSCs) are attractive for cartilage and meniscus regeneration therapy. We developed a software program that can distinguish individual colonies and automatically count the cell number per colony using time-lapse images. In this study, we investigated the usefulness of the software and analyzed colony formation in cultured synovial MSCs. Time-lapse image data were obtained for 14-day-expanded human synovial MSCs. The cell number per colony (for 145 colonies) was automatically counted from phase-contrast and nuclear-stained images. Colony growth curves from day 1 to day 14 (for 140 colonies) were classified using cluster analysis. Correlation analysis of the distribution of the cell number per colony at 14 days versus that number at 1–14 days revealed a correlation at 7 and 14 days. We obtained accurate cell number counts from phase-contrast images. Individual colony growth curves were classified into three main groups and subgroups. Our image analysis software has the potential to improve the evaluation of cell proliferation and to facilitate successful clinical applications using MSCs.
Keywords	Medicine ; R ; Science ; Q
Subject code	571
Language	English
Publishing date	2019-11-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The environmental risk assessment of cell-processing facilities for cell therapy in a Japanese academic institution.

Mitsuru Mizuno / Kentaro Endo / Hisako Katano / Ayako Tsuji / Naomi Kojima / Ken Watanabe / Norio Shimizu / Tomohiro Morio / Ichiro Sekiya

PLoS ONE, Vol 15, Iss 8, p e

2020 Volume 0236600

Abstract: Cell therapy is a promising treatment. One of the key aspects of cell processing products is ensuring sterility of cell-processing facilities (CPFs). The objective of this study was to assess the environmental risk factors inside and outside CPFs. We ... ...

Abstract	Cell therapy is a promising treatment. One of the key aspects of cell processing products is ensuring sterility of cell-processing facilities (CPFs). The objective of this study was to assess the environmental risk factors inside and outside CPFs. We monitored the temperature, humidity, particle number, colony number of microorganisms, bacteria, fungi, and harmful insects in and around our CPF monthly over one year. The temperature in the CPF was constant but the humidity fluctuated depending on the humidity outside. The particle number correlated with the number of entries to the room. Except for winter, colonies of microorganisms and harmful insects were detected depending on the cleanliness of the room. Seven bacterial and two fungal species were identified by PCR analyses. Psocoptera and Acari each accounted for 41% of the total trapped insects. These results provide useful data for taking the appropriate steps to keep entire CPFs clean.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Transplantation of human autologous synovial mesenchymal stem cells with trisomy 7 into the knee joint and 5 years of follow‐up

Mitsuru Mizuno / Kentaro Endo / Hisako Katano / Naoki Amano / Masaki Nomura / Yoshinori Hasegawa / Nobutake Ozeki / Hideyuki Koga / Naoko Takasu / Osamu Ohara / Tomohiro Morio / Ichiro Sekiya

Stem Cells Translational Medicine, Vol 10, Iss 11, Pp 1530-

2021 Volume 1543

Abstract: Abstract Mesenchymal stem cells (MSCs) can show trisomy 7; however, the safety of these cells has not been fully investigated. The purposes of this study were to determine the ratio of patients whose synovial MSCs were transplanted clinically, to ... ...

Abstract	Abstract Mesenchymal stem cells (MSCs) can show trisomy 7; however, the safety of these cells has not been fully investigated. The purposes of this study were to determine the ratio of patients whose synovial MSCs were transplanted clinically, to intensively investigate MSCs with trisomy 7 from a safety perspective, and to follow up the patients for 5 years after transplantation. Synovial MSCs at passage 0 were transplanted into a knee for degenerative meniscus tears in 10 patients, and the patients were checked at 5 years. The synovial MSCs were evaluated at passages 0 to 15 by G‐bands and digital karyotyping, and trisomy 7 was found in 3 of 10 patients. In those three patients, 5% to 10% of the synovial MSCs showed trisomy 7. The mRNA expressions of representative oncogenes and genes on chromosome 7 did not differ between MSCs with and without trisomy 7. Whole‐genome sequencing and DNA methylation analysis showed similar results for MSCs with and without trisomy 7. Transplantation of human synovial MSCs with trisomy 7 into eight mouse knees did not result in tumor formation under the skin or in the knees after 8 weeks in any mouse, whereas transplanted HT1080 cells formed tumors. In vitro chondrogenic potentials were similar between MSCs with and without trisomy 7. Five‐year follow‐ups revealed no serious adverse events in all 10 human patients, including 3 who had received MSCs with trisomy 7. Overall, our findings indicated that synovial MSCs with trisomy 7 were comparable with MSCs without trisomy 7 from a safety perspective.
Keywords	adult stem cells ; cell culture ; clinical trials ; clinical translation ; mesenchymal stem cells (MSCs) ; stem cell culture ; Medicine (General) ; R5-920 ; Cytology ; QH573-671
Subject code	616
Language	English
Publishing date	2021-11-01T00:00:00Z
Publisher	Oxford University Press
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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