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Chondrocyte flow cytometry

Chun-Yuan Chen, Shan-Shan Rao, Lu Ren, Xiong-Ke Hu, Yi-Juan Tan, Yin Hu, Juan Luo, Yi-Wei Liu, Hao Yin, Jie Huang, Jia Cao, Zhen-Xing Wang, Zheng-Zhao Liu, Hao-Ming Liu, Si-Yuan Tang, Ran Xu, Hui Xie
Chronic non-healing wounds represent one of the most common complications of diabetes and need advanced treatment strategies. Exosomes are key mediators of cell paracrine action and can be directly utilized as therapeutic agents for tissue repair and regeneration. Here, we explored the effects of exosomes from human urine-derived stem cells (USC-Exos) on diabetic wound healing and the underlying mechanism. Methods: USCs were characterized by flow cytometry and multipotent differentiation potential analyses...
2018: Theranostics
Ruhang Tang, Liufang Jing, Vincent P Willard, Chia-Lung Wu, Farshid Guilak, Jun Chen, Lori A Setton
BACKGROUND: Intervertebral disc (IVD) degeneration is characterized by an early decrease in cellularity of the nucleus pulposus (NP) region, and associated extracellular matrix changes, reduced hydration, and progressive degeneration. Cell-based IVD therapy has emerged as an area of great interest, with studies reporting regenerative potential for many cell sources, including autologous or allogeneic chondrocytes, primary IVD cells, and stem cells. Few approaches, however, have clear strategies to promote the NP phenotype, in part due to a limited knowledge of the defined markers and differentiation protocols for this lineage...
March 9, 2018: Stem Cell Research & Therapy
Wenhua Pei, Tengfei Lu, Kunfu Wang, Meng Ji, Shuang Zhang, Fenghao Chen, Lu Li, Xiangchen Li, Weijun Guan
Mesenchymal stem cells derived from amniotic fluid have become one of the most potential stem cell source for cell-based therapy for the reason they can be harvested at low cost and without ethical problems. Here, we obtained amniotic fluid stem cells (AFSCs) from ovine amniotic fluid and studied the expansion capacity, cell markers expression, karyotype, and multilineage differentiation ability. In our work, AFSCs were subcultured to passage 62. The cell markers, CD29, CD44, CD73 and OCT4 which analyzed by RT-PCR were positive; CD44, CD73, CD90, CD105, NANOG, OCT4 analyzed by immunofluorescence and flow cytometry were also positive...
March 3, 2018: Cytotechnology
Yiqun Bian, Hao Wang, Shui Sun
Osteoarthritis (OA), characterized by pain and stiffness, swelling, deformity and dysfunction of joints, affects large numbers of population. The purpose of this study was to discover the effects of taurine in human OA chondrocytes and explore the underlying mechanisms. 46 patients with different grades of OA were recruited. Of these patients, 24 underwent total knee replacement and cartilages were harvested. The mRNA expressions of type II collagen (Collagen II) and endoplasmic reticulum (ER) stress markers (GRP78, GADD153 and Caspase-12) in cartilages were quantified by qRT-PCR...
December 2018: Redox Report: Communications in Free Radical Research
Xiong Liu, Yuqi She, Hongrong Wu, Da Zhong, Jian Zhang
BACKGROUND: LncRNA Gas5 is known to be a key control element during growth, differentiation and development in mammalian species. However, the role and function of Gas5 in growth plate chondrocytes has not been determined. METHODS: The overexpression and knockdown models of Gas5 and miR-21 in cells and animals were constructed. Cell survival was determined by MTT assay and flow cytometry. Animal biochemical indices were measured by enzyme-linked immunosorbent assay, hematoxylin/eosin staining, immunohistochemistry or in situ hybridisation...
February 28, 2018: Journal of Biomedical Science
Peng-Fei Hu, Wei-Ping Chen, Jia-Peng Bao, Li-Dong Wu
Apoptosis serves a pivotal role in the pathogenesis of osteoarthritis (OA). Increasing evidence has demonstrated that paeoniflorin exerts key properties (including anticancer, anti-inflammation and neuroprotective) for clinical applications. However, the precise role of paeoniflorin in articular cartilage apoptosis remains unknown. The present study explored the effects and potential molecular mechanism of paeoniflorin on rat chondrocyte apoptosis. Rat articular chondrocytes were cultured in monolayers. The lactate dehydrogenase (LDH) release rate of cells was determined by an LDH release assay...
February 26, 2018: Molecular Medicine Reports
Gangliang Wang, Kangmao Huang, Yangxin Dong, Shuai Chen, Jianfeng Zhang, Jiying Wang, Ziang Xie, Xianfeng Lin, Xiangqiang Fang, Shunwu Fan
BACKGROUND/AIMS: Cartilaginous endplate (CEP) degeneration is an important cause for intervertebral disc (IVD) degeneration that leads to low-back pain. The identification of compounds that may prevent CEP degeneration is of interest for the prevention of IVD degeneration. METHODS: Catabolic protease expression in the CEP of disc degeneration patients was first assessed. The toxicity, function and underlying mechanism of lycorine (LY) on CEP-derived chondrocytes degeneration were assessed in vitro by flow cytometry analysis and western blotting...
February 9, 2018: Cellular Physiology and Biochemistry
Alejandro Prados, Raquel Muñoz-Fernández, Pablo Fernandez-Rubio, Enrique G Olivares
Lymphoid tissue organizer (LTo) cells, identified in mouse and human embryos, are thought to be precursors of stromal cells in secondary lymphoid organs. Whether LTo cells are present in human adults, however remains unknown. We obtained 15 stromal cell lines from tonsils from children who underwent tonsillectomy, and studied the antigen phenotype of these tonsil stromal cell (TSC) lines by flow cytometry and RT-PCR. Cell lines met the minimal criteria proposed by the International Society for Cellular Therapy to define human mesenchymal stem/stromal cells (MSCs): plastic-adherent capacity; expression of CD73, CD90 and CD105, lack of CD45, CD19 and HLA-DR; and capacity to differentiate into adipocytes, osteoblasts and chondrocytes...
February 12, 2018: European Journal of Immunology
Ang Deng, Hongqi Zhang, Minyu Hu, Shaohua Liu, Yuxiang Wang, Qile Gao, Chaofeng Guo
The proliferative rate of chondrocytes affects bone elongation. Chondrocyte hypertrophy is required for endochondral bone formation as chondrocytes secrete factors required for osteoblast differentiation and maturation. Previous studies have demonstrated that the Indian hedgehog (Ihh) signaling pathway is a key regulator of skeletal development and homeostasis. The aim of the present study was to investigate the function of Ihh in chondrocyte proliferation and differentiation, as well as the underlying mechanisms...
January 2018: Experimental and Therapeutic Medicine
Xi-Bin Kao, Qun Chen, Yan Gao, Pin Fan, Jing-Hong Chen, Zhi-Lun Wang, Yan-Qi Wang, Ya-Ni Chen, Yong-Ping Yan
NO plays a key role in the pathological mechanisms of articular diseases. As cytoskeletal proteins are responsible for the polymerization, stabilization, and dynamics of the cytoskeleton network, we investigated whether cytoskeletal proteins are the intracellular pathological targets of NO. We aimed at clarifying whether the cytoskeleton perturbations involved in apoptosis are induced in rabbit articular chondrocytes by NO, which can be liberated by sodium nitroprusside (SNP) treatment. The first passage rabbit articular chondrocytes were cultured as monolayer for the experiments, and the effects of NO were tested in the presence of JNK-specific inhibitor, SP600125...
February 6, 2018: European Journal of Pharmacology
Qi Zhang, Zong-Sheng Yin, Fu-Wen Zhang, Kun Cao, He-Yan Sun
Osteoarthritis (OA), also known as degenerative joint disease or degenerative arthritis, is characterized by chondrocyte apoptosis. The aim of the present study was to investigate the effects of collagen triple helix repeat containing 1 (CTHRC1) and the c‑Jun N‑terminal kinase (JNK) 1/2 inhibitor SP600125 on rat chondrocytes cultured in vitro with interleukin (IL)‑1β. Chondrocytes were treated with different doses of IL‑1β and cell viability and CTHRC1 expression were assessed using Cell Counting Kit‑8 and western blot assays, respectively...
January 18, 2018: International Journal of Molecular Medicine
Junzhong Luo, Jiuhui Han, Yazhou Li, Yuchang Liu
Legg-Calvé-Perthes disease (LCPD) commonly onsets in adolescents, and threatens their health. However, the potential mechanism underlying LCPD remains unclear. MicroRNA (miR)-206 and SRY-box 9 (SOX9) serve an important role in chondrocytes; however, their role in LCPD remains ambiguous. In the present study, whether miR-206 and SOX9 mediated cell apoptosis in dexamethasone (DEX)-induced LCPD was investigated. The chondrocytes of the LCPD and normal control group were isolated from clinical tissues. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the expression of miR-206 and SOX9 mRNA...
January 2018: Oncology Letters
Hori Ghaneialvar, Leila Soltani, Hamid Reza Rahmani, Abbas Sahebghadam Lotfi, Masoud Soleimani
Mesenchymal stem cells are multipotent cells capable of replicating as undifferentiated cells, and have the potential of differentiating into mesenchymal tissue lineages such as osteocytes, adipocytes and chondrocytes. Such lineages can then be used in cell therapy. The aim of present study was to characterize bone marrow derived mesenchymal stem cells in four different species, including: sheep, goat, human and mouse. Human bone-marrow mesenchymal stem cells were purchased, those of sheep and goat were isolated from fetal bone marrow, and those of mouse were collected by washing bone cavity of femur and tibia with DMEM/F12...
January 2018: Indian Journal of Clinical Biochemistry: IJCB
Xiaobo Yang, Weiping Chen, Xiang Zhao, Linwei Chen, Wanli Li, Jisheng Ran, Lidong Wu
Pyruvate kinase M2 (PKM2) has been wildly verified to modulate glycolysis in tumor cells. However, the role of PKM2 on the glycolysis of osteoarthritis (OA) chondrocytes is still unclear. In present study, we investigate the function of PKM2 on OA chondrocyte glycolysis and the collagen matrix generation in vitro. Results showed that PKM2 was upregulated in OA chondrocytes compared with healthy control chondrocytes. In OA chondrocytes, ATP expression was lower compared with healthy control chondrocytes. Loss-of-function experiment showed that PKM2 knockdown mediated by lentivirus transfection could significantly suppress the glucose consumption and lactate secretion levels and decrease glucose transporter-1 (Glut-1), lactate dehydrogenase A (LDHA), and hypoxia inducible factor 1-alpha (HIF-1α), indicating the inhibition of PKM2 knockdown on glycolysis...
January 22, 2018: DNA and Cell Biology
Yongming Xu, Shujuan Xu, Zhi Gao, Liang Xiao, Fei Xiao, Hongguang Xu, Xiaoling Zhang
The aim of this study was to explore the degree of degeneration of endplate chondrocytes in different tension regions induced by intermittent cyclic mechanical tension (ICMT) in vitro. Rat endplate chondrocytes were harvested and treated with 10% ICMT for 8 h/day with a frequency of 0.5 Hz. A cartilage degeneration model was induced using an FX‑5000T cell strain‑loading system. The experiment was divided into the central region and the peripheral region, according to the contact area between the loading post and the six‑well flexible silicone rubber BioFlex plates...
January 16, 2018: Molecular Medicine Reports
Xirui Yang, Qi Zhang, Zhaomeng Gao, Chunyan Yu, Lei Zhang
Baicalin is a flavonoid extracted from Scutellaria baicalensis Georgi, with anti-inflammatory and anti-apoptotic activities. The objective of this study was to explore the effect and mechanism of baicalin on chondrocyte inflammatory response in OA. Different concentrations of IL-1β (0, 0.1, 2, 5 and 10 ng/mL) were used to simulate inflammatory injury in CHON-001 cells. The expression of miR-126 was altered by transfection with miR-126 mimic. Thereafter, cells were treated with baicalin, and cell viability, apoptosis, the expressions of apoptosis-related protein and pro-inflammatory factors were respectively detected using CCK-8 assay, flow cytometry, qRT-PCR and western blot analysis...
January 11, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Hao Wang, Xu Yan, Yuxin Jiang, Zheng Wang, Yufei Li, Qingdong Shao
Osteoarthritis (OA) affects a large number of patients; however, human umbilical cord stem cells exhibit therapeutic potential for treating OA. The aim of the present study was to explore the interaction between human umbilical cord stem cells and degenerated chondrocytes, and the therapeutic potential of human umbilical cord stem cells on degenerated chondrocytes. Human umbilical cord‑derived mesenchymal stem cells (hUC‑MSCs) were harvested from human umbilical cords, and flow cytometry was used to analyze the surface antigen markers, in addition, chondrogenic, osteogenic and adipogenic differentiation on the cells was investigated...
January 9, 2018: Molecular Medicine Reports
Rana Moradian Tehrani, Hamed Mirzaie, Javad Verdi, Amirhossein Sahebkar, Mahdi Noureddini, Rasoul Salehi, Behrang Alani, Mojtaba Kianmehr
Human adipose tissue has been identified as a viable alternative source for mesenchymal stem cells. SADSCs were isolated from human scalp biopsy and then were characterized by Flow cytometry. SADSCS expressed CD90, CD44, and CD105 but did not express CD45 surface marker. Growth factors were used for chondrogenesis induction. Histology and immunohistology methods and gene expression by real-time PCR 14 days after induced cells have shown the feature of chondrocytes in their morphology and extracellular matrix in both inducing patterns of combination and cycling induction...
January 11, 2018: Journal of Cellular Physiology
Chuandong Yu, Yongkun Wang
BACKGROUND: Osteoarthritis (OA), as a degenerative disease, is a major problem in ageing populations. To better understand the underlying mechanisms in the pathogenesis of OA, this study was undertaken to investigate the role of microRNA (miR)-19a in chondrocytes. METHODS: Expression of the members of miR-17-92 cluster in synovium from OA patients and non-OA patients were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). miR-19a was abnormal expressed in human chondrocyte line (CHON-001 and T-C/28 cells) and primary human chondrocytes by transient transfection...
January 3, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
You Sun, Feifei Wang, Xuehui Sun, Xiaoxue Wang, Lei Zhang, Yang Li
OBJECTIVE: The study was aimed to investigate the impact of CX3CR1 expression on the proliferation and apoptosis of osteoarthrosis (OA) chondrocytes through Wnt/ β-catenin pathway. METHODS: The expression levels of CX3CR1 and Wnt/β-catenin pathway-related genes mRNA and protein in OA chondrocytes were examined by qRT-PCR and western blot. MTT and flow cytometry (FCM) assays were employed to assess cell proliferation, cell cycle and apoptosis. XAV-939, a Wnt/β-catenin pathway inhibitor, was used to inhibit the pathway...
December 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
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