Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts

Casnici, Claudia; Lattuada, D.; Tonna, Noemi; Crotta, Katia; Storini, Claudio; Bianco, Fabio; Truzzi, Marcello; Corradini, C.; Marelli, O

doi:10.1155/2014/702057

Cited by 21 publications

(17 citation statements)

References 35 publications

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“…Synovial membranes were harvested during arthroplasty from an RA patient at our institution with active inflammation who provided written informed consent to participate in the study. Primary RA synovial fibroblasts were isolated based on a previous procedure that describes the successful isolation of human RA synovial fibroblasts with minor modifications 43 . Briefly, synovial tissues from RA patients were stored in 70% ethanol at the operation room; in the laboratory, the samples were minced and incubated for 4 h with 2.5 mg/mL type I collagenase (Sigma Aldrich, St. Louis, MO, USA) in Roswell Park Memorial Institute (RPMI) 1640 (Sigma-Aldrich) at 37 °C in 5% CO 2 .…”

Section: Harvesting Of Adipose Tissue and Adipose Stem Cell Isolationmentioning

confidence: 99%

Local transplantation of adipose-derived stem cells has a significant therapeutic effect in a mouse model of rheumatoid arthritis

Ueyama

Okano

Orita

et al. 2020

Sci Rep

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Adipose-derived stem cells (ADSCs) have anti-inflammatory and regenerative properties. The purpose of this study was to investigate the effect of locally administered ADSCs in a rheumatoid arthritis (RA) mouse model. In an in vivo experiment, single-cell ADScs and three dimensionally-cultured ADSc spheroids were injected intra-articularly into the knees of RA model mice and histologically assessed. Marked improvement of synovial inflammation and articular cartilage regeneration was found in ADSCtreated mice. Proliferation, migration, and apoptosis assays of synovial fibroblasts incubated with single-cell and spheroid ADSCs were performed. The expression levels of total cytokine RNA in ADSC single cells, spheroids, and ADSC-treated inflammatory synovial fibroblasts were also evaluated by quantitative reverse transcription PCR. ADSCs suppressed the proliferation and migration of activated inflammatory cells and downregulated inflammatory cytokines. TSG-6 and TGFβ1 were significantly upregulated in ADScs compared to controls and TGFβ1 was significantly upregulated in ADSC spheroids compared to single cells. The apoptosis rate of ADSC spheroids was significantly lower than that of single-cell ADSCs. These results indicated that intra-articular administration of ADSC single cells and spheroids was effective in an RA mouse model, offering a novel approach for the development of effective localized treatments for patients with RA. Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovial inflammation and degeneration of articular cartilage and bone. In previous decades, systemic therapies, including methotrexate and biological disease-modifying antirheumatic drugs, significantly improved the treatment of synovial inflammation 1. Given that RA is a systemic disease, the focus has been to develop systemic treatments for it 2 ; however, swelling persists in some joints with such treatments and the application of systemic treatment is limited due to adverse, off-target effects. Therefore, localized treatment methods are important for treating RA. Corticosteroid injections are the most widely used localized treatment for RA. Although it reduces RA-associated synovitis, repeated intra-articular corticosteroid injections can lead to the degeneration of articular cartilage 3. Other adverse effects of corticosteroids include septic arthritis, local tissue atrophy, tendon rupture, and hyperglycaemia. Consequently, the development of safe, effective, and tissue-regenerating localized therapies must be explored. Recent studies in regenerative medicine have used mesenchymal stem cells (MSCs) derived from somatic tissues such as bone marrow, muscle, blood, and adipose tissue 4,5. MSCs exert anti-inflammatory effects, modulate the immune response, and modify the local microenvironment; furthermore, studies have reported their therapeutic effects for the treatment of inflammatory and autoimmune diseases 6,7. Considering the accessibility and ethical issues associated with the use of MSCs, adipose-derived stem cells (...

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Section: Harvesting Of Adipose Tissue and Adipose Stem Cell Isolationmentioning

confidence: 99%

Local transplantation of adipose-derived stem cells has a significant therapeutic effect in a mouse model of rheumatoid arthritis

Ueyama

Okano

Orita

et al. 2020

Sci Rep

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“…In the other method, the cells are plated at high density. The cells obtained by plating at high density are referred to as synovial cells, synovial fibroblasts, and synoviocytes . We hypothesized that the initial cell plating density affected the properties of primary synovial MSCs.…”

mentioning

confidence: 99%

“…The cells obtained by plating at high density are referred to as synovial cells, synovial fibroblasts, and synoviocytes. 10,11 We hypothesized that the initial cell plating density affected the properties of primary synovial MSCs. Hence, the purpose of this study is to investigate the effects of initial cell density on proliferation, surface markers, colony-forming ability, and multipotentiality, including chondrogenesis in synovial MSCs.…”

mentioning

confidence: 99%

Initial cell plating density affects properties of human primary synovial mesenchymal stem cells

Nakamura¹,

Tsuji

Mizuno

et al. 2018

Journal Orthopaedic Research

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Synovial mesenchymal stem cells (MSCs) appear to be an attractive cell source in cartilage and meniscus regeneration because of their high proliferative and chondrogenic potentials. Two methods are used to culture synovial nucleated cells in the preparation of primary synovial MSCs. In one method, the cells are plated at low density to make cell colonies. In the other method, the cells are plated at high density. We investigated the effects of initial cell density on proliferation, surface markers, and multipotentiality, including chondrogenesis in primary synovial MSCs. Human synovium was obtained from the knee joints of patients with osteoarthritis after total knee arthroplasty. Immediately after enzyme digestion, the synovial nucleated cells were plated in densities of 10 , 10 , or 10 cells/60-cm dish and cultured for 14 days. Proliferation, surface markers, chondrogenesis, adipogenesis, and calcification were examined in three populations. The cell colonies were distinct in the 10 cells/dish group, faint in the 10 cells/dish group, and obscure in the 10 cells/dish group. The total number of cells/dish was positively related to plating density, whereas the fold increase was negatively related to plating density (n = 13). Among 12 surface markers, a negative relation to plating density was distinct in CD105. The cartilage pellet weight was negatively related to the initial plating density. The oil red-o positive area and alizarin red positive area were positively related to the initial plating density. The initial cell plating density affected the properties of primary synovial MSCs. Synovial nucleated cells proliferated better when plated at low density, and the synovial MSCs obtained by this method contained a high chondrogenic potential. © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. J Orthop Res.

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“…In agreement with these results, Lattuada et al (2016) proved that the percentage of apoptotic cells (RA-FLSs) cultured in synovial fluid samples aspirated from the joints of RA was lower than in cells cultured in tissue medium alone, probably because of the presence in the SF of anti-apoptotic factors. Resistance to apoptosis in RA-FLSs depends on up regulation of IAPs which are expressed at high levels in RA-FLSs (Casnici et al, 2014). Also, Jia et al (2015) detected increased proliferation and inhibited apoptosis in synovial cells separated from synovial tissues harvested from RA patients.…”

Section: Discussionmentioning

confidence: 93%

Evaluation of the effect of vitamin C on caspase 9 and oxidative stress in rheumatoid arthritis patients

Elshamy¹,

Gaafar²,

ElAshwah³

et al. 2018

Afr. J. Biochem. Res.

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Rheumatoid arthritis (RA) is a chronic and an autoimmune disease of the joints and is widely distributed worldwide. It is characterized by alterations of the antioxidant defense system and increased free radical formation and pro-inflammatory cytokine. The aim of the present study was to evaluate the effect of vitamin C supplementation on oxidative stress biomarkers and caspase 9 level in rheumatoid arthritis patients. This study included 30 RA patients and 30 healthy subjects. Plasma levels of malondialdehyde (MDA), total antioxidant capacity (TAC), caspase 9 and 8-hydroxy-2′deoxyguanosine (8 OHdG) were assayed as well as blood vitamin C level. These parameters were reevaluated in RA patients after vitamin C supplementation for one month. Increased MDA and 8 OHdG levels and reduced TAC, caspase 9 and vitamin C. Levels were demonstrated in RA patients. After vitamin C supplementation, RA patients showed significant increase in TAC and vitamin C level and significant decrease in MDA and 8 OHdG levels, plasma caspase 9 level was not significantly affected after vitamin C supplementation. Increased oxidative stress and decreased apoptosis may have an important role in the pathogenesis of RA. The administration of vitamin C supplementation may help to relieve oxidative stress and enhance the antioxidant defense in these patients.

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Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts

Cited by 21 publications

References 35 publications

Local transplantation of adipose-derived stem cells has a significant therapeutic effect in a mouse model of rheumatoid arthritis

Local transplantation of adipose-derived stem cells has a significant therapeutic effect in a mouse model of rheumatoid arthritis

Initial cell plating density affects properties of human primary synovial mesenchymal stem cells

Evaluation of the effect of vitamin C on caspase 9 and oxidative stress in rheumatoid arthritis patients

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