Analysis of the chondrogenic potential of human synovial stem cells according to harvest site and culture parameters in knees with medial compartment osteoarthritis

Nagase, Tsuyoshi; Muneta, Takeshi; Ju, Young-Jin; Hara, Kenji; Morito, Toshiyuki; Koga, Hideyuki; Nimura, Akimoto; Mochizuki, Takashi; Sekiya, Ichiro

doi:10.1002/art.23418

Cited by 94 publications

(84 citation statements)

References 41 publications

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“…It is suggested that neo-cartilage in 3D culture is observed after 14-21 days of continuous culture. Together with the previous literature, this refl ected the fact that the duration of our 3D culture was too short to allow re-differentiation from the monolayer culture of ACs and SYs passaged continuously (Nagase et al, 2008;Zhang et al, 2004). Furthermore, our system can yield mass production of transplantable chondrogenic tissue.…”

Section: Spheroids Of Synovium-derived Cells and Chondrocytessupporting

confidence: 54%

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Transplantation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee

Lee

Sato²,

Kim³

et al. 2011

eCM

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Autologous chondrocyte implantation (ACI) is the treatment of choice for osteoarthritis. However, to regenerate articular cartilage using this method, the procedure paradoxically demands that the cell source of the articular chondrocytes (ACs) for ex vivo expansion be from the patient's own healthy cartilage, which can result in donor site morbidity. Accordingly, it is essential to develop a substitute for AC. In the present study, we investigated whether synovium-derived cells (SYs) could be used as a partial replacement for ACs in ACI. ACs and SYs from the knees of rabbits were isolated and cultured, and the growth rates of the cells were compared. To manufacture the cellular transplants, we developed a high-density suspension-shaking culture method (HDSS), which circulates the cells in culture media, promoting self-assembly of scaffold-free cellular aggregates. ACs and SYs were mixed in various ratios using HDSS. Injectable cellular transplants were harvested and transplanted into full-thickness osteochondral defects. Simultaneously, histological evaluations were conducted with toluidine blue and safranin O, and immunohistochemistry of collagen type I and II was conducted. Gene expression to evaluate chondrocyte-specifi c differentiation was also performed. We successfully prepared a large quantity of spheroids (spheroidal cell aggregates) in a short time using mixed ACs and SYs, for all cellular composition ratios. Our data showed that the minimal therapeutic unit for the transplants contributed to in situ regeneration of cartilage. In summary, SYs can be used as a replacement for ACs in clinical cases of ACI in patients with broad areas of osteoarthritic lesions.

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Section: Spheroids Of Synovium-derived Cells and Chondrocytessupporting

confidence: 54%

“…their high proliferative capacity and chondrogenic potential ( Sakaguchi et al, 2005;Yoshimura et al, 2007;Nagase et al, 2008;.…”

Section: Spheroids Of Synovium-derived Cells and Chondrocytesmentioning

confidence: 99%

Transplantation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee

Lee

Sato²,

Kim³

et al. 2011

eCM

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“…Cell fluorescence was evaluated by FACSCalibur instrument (Becton Dickinson), and data were analyzed using CellQuest software (Becton Dickinson). 11,15 Chondrogenesis Two hundred thousands cells at passage 2 were placed in a 15-ml polypropylene tube (Becton Dickinson) and centrifuged at 450 g for 10 min. The pellets were cultured at 371C with 5% CO 2 in 400 ml chondrogenic media that contained 500 ng/ml bone morphogenetic protein 2 (Astellas Pharm, Tokyo, Japan), 10 ng/ml transforming growth factor-b3 (R&D Systems, Minneapolis, MN, USA), 100 nM dexamethasone, 50 mg/ml ascorbate-2-phosphate, 40 mg/ml proline, 100 mg/ml pyruvate (Sigma-Aldrich), and 50 mg/ml ITS þ Premix in high-glucose Dulbecco modified Eagle medium (Invitrogen).…”

Section: Surface Epitopesmentioning

confidence: 99%

“…The medium was replaced every 3-4 days for 21 days. [15][16][17][18] Histology The cultures were ended by fixing the pellets with 2.5% glutaraldehyde in 0.1 M PBS for 2 h. The cells were washed overnight at 41C in the same buffer and post-fixed with 1% OsO4 buffered with 0.1 M PBS for 2 h. The pellets were dehydrated in a graded series of ethanol and embedded in Epon 812. Semi-thin (1 mm) sections for light microscopy were collected on glass slides and stained for 30 s with toluidine blue.…”

Section: Surface Epitopesmentioning

confidence: 99%

Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes

Ichinose

Muneta

Koga

et al. 2010

Laboratory Investigation

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Mesenchymal stem cells (MSCs) from a variety of mesenchymal tissue contain common features, but distinguishing properties dependent on their origin are emerging. We investigated morphological differences of human bone marrowMSCs, synovium-MSCs, and chondrocytes during in vitro chondrogenesis. Two hundred thousands cells were pelleted after centrifugation and cultured in chondrogenic media that contained BMP-2, TGF-b3, and dexamethasone. The pellets were analyzed histologically, immunohistologically, and electron microscopically. Before chondrogenic induction, trypsinized MSCs and chondrocytes looked similar. At day 1, the structure of the three masses was divided into two layers, and the most obvious differences in the three populations were observed in the deep zone. In bone marrow-MSCs, round cells accumulated without intercellular space, and the cells were mainly connected through intermediate junctions. In synovium-MSCs, elongated cells accumulated with small desmosomes and intercellular spaces could occasionally be seen. In chondrocytes, separated oval and polygonal cells connected only in a narrow spotty area through a small desmosome. At day 7, the structure of the three masses was divided into three layers, and the most obvious differences in the three populations were observed in the middle zone. In bone marrow-MSCs, the middle zone consisted of dense smaller cells and apoptotic cells. In synovium-MSCs, the middle zone consisted of dense arrayed wider cells and apoptotic cells. In chondrocytes, the middle zone was acellular without apoptotic cells. At day 21, the morphology of cells and extracellular space became similar in that each cell was located separately with abundant extracellular matrix. The superficial zone was still obvious in bone marrow-MSCs, but hardly seen both in synovium-MSCs and chondrocytes. In this study, we revealed morphological differences of bone marrow-MSCs, synovium-MSCs, and chondrocytes during in vitro chondrogenesis. The most obvious differences in the three populations were observed at day 1 in the deep zone.

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“…SDSCs maintain their chondrogenic capacity regardless of donor age or disease condition (De Bari et al, 2001;Nagase et al, 2008) and have potential for tissue-engineering applications aimed at cartilage repair or regeneration (Bilgen et al, 2007;Han et al, 2010;Lima et al, 2007;Pei et al, 2008a;Pei et al, 2008b). We previously cultured SDSCs in a 3D hydrogel which achieved properties approaching those of native juvenile bovine cartilage.…”

Section: Introductionmentioning

confidence: 99%

Applied Osmotic Loading for Promoting Development of Engineered Cartilage

Sampat

Robinson

Ackerman

et al. 2012

ASME 2012 Summer Bioengineering Conference, Parts a and B

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This study investigated the potential use of static osmotic loading as a cartilage tissue engineering strategy for growing clinically relevant grafts from either synovium-derived stem cells (SDSCs) or chondrocytes. Bovine SDSCs and chondrocytes were individually encapsulated in 2% w/v agarose and divided into chondrogenic media of osmolarities 300 (hypotonic), 330 (isotonic), and 400 (hypertonic, physiologic) mOsM for up to 7 weeks. The application of hypertonic media to constructs comprised of SDSCs or chondrocytes led to increased mechanical properties as compared to hypotonic (300 mOsM) or isotonic (330 mOsM) media (p<0.05). Constant exposure of SDSC-seeded constructs to 400 mOsM media from day 0 to day 49 yielded a Young's modulus of 513±89 kPa and GAG content of 7.39±0.52% ww on day 49, well within the range of values of native, immature bovine cartilage. Primary chondrocyte-seeded constructs achieved almost as high a Young's modulus, reaching 487±187 kPa and 6.77±0.54%ww (GAG) for the 400 mOsM condition (day 42). These findings suggest hypertonic loading as a straightforward strategy for 3D cultivation with significant benefits for cartilage tissue engineering strategies. In an effort to understand potential mechanisms responsible for the observed response, cell volume measurements in response to varying osmotic conditions were evaluated in relation to the Boylevan't Hoff (BVH) law. Results confirmed that chondrocytes behave as perfect osmometers; however SDSCs deviated from the BVH relation.

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Analysis of the chondrogenic potential of human synovial stem cells according to harvest site and culture parameters in knees with medial compartment osteoarthritis

Cited by 94 publications

References 41 publications

Transplantation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee

Transplantation of scaffold-free spheroids composed of synovium-derived cells and chondrocytes for the treatment of cartilage defects of the knee

Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes

Applied Osmotic Loading for Promoting Development of Engineered Cartilage

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