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Chondrocyte flow cytometry

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https://www.readbyqxmd.com/read/28024508/-reconstruction-of-humanized-bone-marrow-niche-in-immunodeficiency-mouse
#1
Huan Chen, Zheng Tian, Bin Li, Hai-Yan Xing, Huan Li, Ke-Jing Tang, Min Wang, Qing Rao
OBJECTIVE: To reconstruct a human bone marrow niche in immunodeficiency mouse (NOD/SCID) so as to provide a model for observing the effect of abnormal BM niche on the occurence and development of leukemia. METHODS: Human platelet lysate(HPL) was obtained by repeated freezing and thawing of concentrated platelet. Bone marrow-derived mesenchymal stem cells were cultured in α-minimal essential medium (α-MEM) containing 10% HPL or 10% FBS. The morphology, cell phenotype, multilineage differentiation potential in vitro and proliferation capacity between the mesenchymal stem cells cultured with HPL or FBS were compared...
December 2016: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28002898/tranexamic-acid-effects-on-cartilage-and-synovial-tissue-an-in-vitro-study-for-a-possible-safe-intra-articular-use
#2
A Marmotti, S Mattia, L Mangiavini, D E Bonasia, M Bruzzone, F Dettoni, F Rosso, D Blonna, R Rossi, F Castoldi, G M Peretti
The possible toxic effects of intra-articular tranexamic acid (TA) are still debated. The aim of this study was to evaluate TA effects on human cartilage fragments and synovial biopsies. Explant culture of minced articular cartilage underwent prolonged TA exposure. Histological analysis, immunofluorescence and colorimetric assay for quantification of s-GAG and DNA were performed at the end term. Synoviocytes were cultured for 48h in presence of TA. Light microscopy and flow cytometry analysis were performed at the end of the exposure to TA and one week after the treatment...
October 2016: Journal of Biological Regulators and Homeostatic Agents
https://www.readbyqxmd.com/read/27925477/relevance-of-meniscal-cell-regional-phenotype-to-tissue-engineering
#3
Shawn P Grogan, Chantal Pauli, Martin K Lotz, Darryl D D'Lima
PURPOSE: Meniscus contains heterogeneous populations of cells that have not been fully characterized. Cell phenotype is often lost during culture; however, culture expansion is typically required for tissue engineering. We examined and compared cell surface molecule expression levels on human meniscus cells from the vascular and avascular regions and articular chondrocytes while documenting changes during culture-induced dedifferentiation. MATERIALS AND METHODS: Expression of 16 different surface molecules were examined by flow cytometry after monolayer culture for 24 h, 1 week, and 2 weeks...
December 7, 2016: Connective Tissue Research
https://www.readbyqxmd.com/read/27916780/mir-15a-5p-regulates-viability-and-matrix-degradation-of-human-osteoarthritis-chondrocytes-via-targeting-vegfa
#4
Hongwei Chen, Yun Tian
Previous studies demonstrated that miR-15a-5p was probably associated with human hepatocellular carcinoma, while the function of miR-15a-5p in OA (Osteoarthritis) still remains unknown. Here, we uncovered the potential role of miR-15a-5p on OA pathogenesis and confirmed its predicted target VEGFA (Vascular Endothelial Growth Factor A). Measured by RT-PCR, miR-15a-5p expression increased remarkably while VEGFA expression was significantly decreased in OA chondrocytes compared with normal conditions. According to Luciferase activity assay, miR-15a-5p directly targeted the 3'-UTR of VEGFA to inhibit its expression...
December 3, 2016: Bioscience Trends
https://www.readbyqxmd.com/read/27910811/ll-37-hnp-1-and-hbd2-3-modulate-the-secretion-of-cytokines-tnf-%C3%AE-il-6-ifn-%C3%AE-il-10-and-mmp1-in-human-primary-cell-cultures
#5
Carlos Erik Medina Santos, Carmen Nathaly López Hurtado, Bruno Rivas Santiago, Roberto Gonzalez-Amaro, Yolanda Guadalupe Cataño Cañizales, Margarita de la Luz Martínez Fierro, José Antonio Enciso-Moreno, Mariana Haydee García Hernández
The aim of this study was to evaluate the effects of the LL-37, HNP-1 and HBD2/3 peptides on cytokine and MMP production in human polymorphonuclear cells, mononuclear cells and chondrocytes. The levels of cytokines in supernatants from mononuclear and polymorphonuclear cell cultures were measured with a cytometric bead array by flow cytometry. Likewise, the levels of metalloproteinase/MMP-1, 3, and 13 were measured in supernatants from chondrocyte cultures using an ELISA. The expression of RANKL on lymphocytes was analyzed by flow cytometry...
September 1, 2016: European Cytokine Network
https://www.readbyqxmd.com/read/27907835/inhibition-of-the-p38-mapk-signaling-pathway-suppresses-the-apoptosis-and-expression-of-proinflammatory-cytokines-in-human-osteoarthritis-chondrocytes
#6
He-Yan Sun, Kong-Zu Hu, Zong-Sheng Yin
This study aims to investigate the effects of p38-MAPK signaling pathway on the apoptosis and expression of proinflammatory cytokines in human osteoarthritis (OA) chondrocytes. Human articular cartilage specimens were obtained from 57 OA patients and 31 patients with lower extremity traumatic amputations. The expressions of p38-MAPK pathway-related proteins in cartilage tissue were detected by immunochemistry. Cultured chondrocytes isolated from human OA cartilage were assigned into the blank group, the IL-1β group, the PD (PD980959, ERK pathway inhibitors)+IL-1β group, the SB (SB203580, p38 pathway inhibitors)+IL-1β group, and the SP (SP600125, JNK signaling pathway inhibitors)+IL-1β group...
November 28, 2016: Cytokine
https://www.readbyqxmd.com/read/27882184/phosphorylation-of-osteopontin-has-proapoptotic-and-proinflammatory-effects-on-human-knee-osteoarthritis-chondrocytes
#7
Shu-Guang Gao, Yang Yu, Chao Zeng, Shi-Tao Lu, Jian Tian, Chao Cheng, Liang-Jun Li, Guang-Hua Lei
The aim of the present study was to investigate the effects of phosphorylated osteopontin (p-OPN) on apoptosis and pro-inflammatory cytokine expression in human knee osteoarthritis (OA) chondrocytes. Human knee OA chondrocytes obtained from patients who underwent total knee arthroplasty were treated with p-OPN, OPN or buffer. Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) and western blot analysis were used to assess the expression levels of proinflammatory factors, including interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6 and nuclear factor (NF)-κB...
November 2016: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/27864696/generation-and-differentiation-of-induced-pluripotent-stem-cells-reveal-ankylosing-spondylitis-risk-gene-expression-in-bone-progenitors
#8
Gerlinde Layh-Schmitt, Shajia Lu, Fatemeh Navid, Stephen R Brooks, Emily Lazowick, Kathryn M Davis, Cristina Montagna, Massimo Gadina, Robert A Colbert
Axial spondyloarthritis (axSpA), which encompasses ankylosing spondylitis, is a complex genetic disease. Aberrant bone formation is a key feature of pathogenesis that can lead to ankylosis of the spine. Our objective is to determine, whether genes whose variants confer susceptibility to AS are expressed in bone progenitors like mesenchymal stem cells (MSCs). Since MSCs from bone marrow is difficult to obtain, we first examined, whether MSCs can be derived from induced pluripotent stem cells (iPSCs). Dermal fibroblasts of two axSpA patients and one healthy control were reprogrammed into iPSCs using a Sendai virus vector encoding pluripotency genes...
January 2017: Clinical Rheumatology
https://www.readbyqxmd.com/read/27864279/nutrition-deficiency-promotes-apoptosis-of-cartilage-endplate-stem-cells-in-a-caspase-independent-manner-partially-through-upregulating-bnip3
#9
Zhiliang He, Luqiao Pu, Chao Yuan, Min Jia, Jian Wang
Nutrition deficiency is reported to induce apoptosis of chondrocytes and degeneration of cartilage endplate (CEP) in rabbit. Cartilage endplate stem cells (CESCs) are important for the integrity of structure and function of CEP. Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (BNIP3) has been reported to regulate apoptosis, autophagy, and cytoprotection. In this study, we aimed to determine whether nutrition deficiency induces apoptosis of CESCs, and whether or not the BNIP3-related pathway is activated in CESCs during nutrition deficiency...
January 2017: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/27851782/alpha-b-crystallin-protects-rat-articular-chondrocytes-against-casein-kinase-ii-inhibition-induced-apoptosis
#10
Sung Won Lee, Jee Hyun Rho, Sang Yeob Lee, Seung Hee Yoo, Hye Young Kim, Won Tae Chung, Young Hyun Yoo
Although alpha (α)B-crystallin is expressed in articular chondrocytes, little is known about its role in these cells. Protein kinase casein kinase 2 (CK2) inhibition induces articular chondrocyte death. The present study examines whether αB-crystallin exerts anti-apoptotic activity in articular chondrocytes. Primary rat articular chondrocytes were isolated from knee joint slices. Cells were treated with CK2 inhibitors with or without αB-crystallin siRNA. To examine whether the silencing of αB-crystallin sensitizes rat articular chondrocytes to CK2 inhibition-induced apoptosis, we assessed apoptosis by performing viability assays, mitochondrial membrane potential measurements, flow cytometry, nuclear morphology observations, and western blot analysis...
2016: PloS One
https://www.readbyqxmd.com/read/27778036/-biological-characteristics-of-mesenchymal-stem-cell-and-hematopoietic-stem-cell-in-the-co-culture-system
#11
Wei Wei, Chao Xu, Zhi-Yong Ye, Xiao-Jun Huang, Jia-En Yuan, Tian-Bao Ma, Han-Biao Lin, Xiu-Qiong Chen
The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34(+) HSC was collected by MACS immunomagnetic beads. The selected CD34(+) HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC...
October 25, 2016: Sheng Li Xue Bao: [Acta Physiologica Sinica]
https://www.readbyqxmd.com/read/27702388/epidermal-growth-factor-induces-proliferation-of-hair-follicle-derived-mesenchymal-stem-cells-through-epidermal-growth-factor-receptor-mediated-activation-of-erk-and-akt-signaling-pathways-associated-with-upregulation-of-cyclin-d1-and-downregulation-of-p16
#12
Tingting Bai, Feilin Liu, Fei Zou, Guifang Zhao, Yixu Jiang, Li Liu, Jiahong Shi, Deshun Hao, Qi Zhang, Tong Zheng, Yingyao Zhang, Mingsheng Liu, Shilun Li, Liangchen Qi, Jin Yu Liu
The maintenance of highly proliferative capacity and full differentiation potential is a necessary step in the initiation of stem cell-based regenerative medicine. Our recent study showed that epidermal growth factor (EGF) significantly enhanced hair follicle-derived mesenchymal stem cell (HF-MSC) proliferation while maintaining the multilineage differentiation potentials. However, the underlying mechanism remains unclear. Herein, we investigated the role of EGF in HF-MSC proliferation. HF-MSCs were isolated and cultured with or without EGF...
January 15, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/27665780/platelet-rich-plasma-protects-rat-chondrocytes-from-interleukin-1%C3%AE-induced-apoptosis
#13
Jinjiang Yang, Ying Lu, Ai Guo
Interleukin (IL)-1β-induced chondrocyte apoptosis is associated with the pathogenesis of arthritis. Platelet‑rich plasma (PRP), which is derived from the patient's own blood and contains numerous growth factors, has the potential for arthritis treatment. Therefore, the present study aimed to determine the effects of PRP on chondrocyte apoptosis, under IL‑1β‑induced pathological conditions. Chondrocytes isolated from the knee joint of Sprague Dawley rats were used in the present study. Cell viability was determined using the Cell Counting kit‑8 assay, cell apoptosis was evaluated by flow cytometry, and the expression of apoptosis‑, anabolism‑ and catabolism-associated genes were detected by quantitative polymerase chain reaction; protein expression was detected by western blot analysis...
November 2016: Molecular Medicine Reports
https://www.readbyqxmd.com/read/27609477/comparative-characteristics-of-mesenchymal-stem-cells-derived-from-reamer-irrigator-aspirator-iliac-crest-bone-marrow-and-adipose-tissue
#14
S Toosi, H Naderi-Meshkin, F Kalalinia, M T Peivandi, H Hossein Khani, A R Bahrami, A Heirani-Tabasi, M Mirahmadi, J Behravan
Mesenchymal stem cells (MSCs) have been considered promising tools for new clinical concepts in supporting cellular therapy and regenerative medicine. More recently, Ream/Irrigator/Aspirator (RIA) was introduced as a source of MSCs. In this study we compared MSCs derived from three different sources (iliac crest bone marrow (ICBM), adipose tissue (AT), and (RIA)) regarding the morphology, the success rate of isolating MSCs, colony frequency, expansion potential, osteogenic and chondrogenic differentiation capacity...
August 31, 2016: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/27574269/alloreactivity-and-immunosuppressive-properties-of-articular-chondrocytes-from-osteoarthritic-cartilage
#15
S Abe, H Nochi, H Ito
PURPOSE: To determine whether articular chondrocytes derived from osteoarthritic knee joints could evoke alloreactive proliferation of peripheral blood mononuclear cells (PBMC) and inhibit mitogenic activity of polyclonally activated CD4+ major histocompatibility complex (MHC) class II- restricted T cells in vitro. METHODS: Osteoarthritic cartilages of 17 patients aged 61 to 85 years were harvested during total knee arthroplasty. Chondrocytes were cultured for experiments...
August 2016: Journal of Orthopaedic Surgery
https://www.readbyqxmd.com/read/27553254/differentiation-potential-of-rabbit-cd90-positive-cells-sorted-from-adipose-derived-stem-cells-in-vitro
#16
Xinghui Song, Chaoyang Hong, Qingqing Zheng, Hailan Zhao, Kangping Song, Zhe Liu, Jiang Shen, Yanwei Li, Jiajia Wang, Ting Shen
To investigate the differentiation potential of purified CD90(+) cells sorted from adipose-derived stem cells (ADSCs), CD90(+) cells were sorted from rabbit ADSCs using flow cytometry. Then, cell expansion of CD90(+) cells and unsorted ADSCs was observed using an inverted microscope. Furthermore, cell surface markers including CD40, CD105, and CD90 on CD90(+) cells and unsorted ADSCs were quantified using flow cytometry. Additionally, multi-lineage differentiation ability between CD90(+) cells and unsorted ADSCs was compared, and expression of adipocyte-related genes PPAR-r and CEBPA as well as stem cell-related gene SOX2 in CD90(+) cells and unsorted ADSCs was determined using real-time quantitative PCR...
August 23, 2016: In Vitro Cellular & Developmental Biology. Animal
https://www.readbyqxmd.com/read/27495937/rapid-generation-of-functional-hepatocyte-like-cells-from-human-adipose-derived-stem-cells
#17
Yanli Fu, Jie Deng, Qingyuan Jiang, Yuan Wang, Yujing Zhang, Yunqi Yao, Fuyi Cheng, Xiaolei Chen, Fen Xu, Meijuan Huang, Yang Yang, Shuang Zhang, Dechao Yu, Robert Chunhua Zhao, Yuquan Wei, Hongxin Deng
BACKGROUND: Liver disease is a major cause of death worldwide. Orthotropic liver transplantation (OLT) represents the only effective treatment for patients with liver failure, but the increasing demand for organs is unfortunately so great that its application is limited. Hepatocyte transplantation is a promising alternative to OLT for the treatment of some liver-based metabolic disorders or acute liver failure. Unfortunately, the lack of donor livers also makes it difficult to obtain enough viable hepatocytes for hepatocyte-based therapies...
August 5, 2016: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/27489592/human-platelet-lysate-as-a-xeno-free-alternative-of-fetal-bovine-serum-for-the-in-vitro-expansion-of-human-mesenchymal-stromal-cells
#18
Saeed Mohammadi, Mohsen Nikbakht, Ashraf Malek Mohammadi, Mahdi Zahed Panah, Mohammad Reza Ostadali, Hajar Nasiri, Ardeshir Ghavamzadeh
BACKGROUND: Mesenchymal stromal cells (MSCs) are employed in various different clinical settings in order to modulate immune response. Human autologous and allogeneic supplements including platelet derivatives such as platelet lysate (PL), platelet-released factors (PRF) and serum are assessed in clinical studies to replace fetal bovine serum (FBS). The immunosuppressive activity and multi-potential characteristic of MSCs appear to be maintained when the cells are expanded in platelet derivatives...
July 1, 2016: International Journal of Hematology-oncology and Stem Cell Research
https://www.readbyqxmd.com/read/27466293/oleic-palmitate-induces-apoptosis-in-human-articular-chondrocytes-via-upregulation-of-nox4-expression-and-ros-production
#19
Dapeng Fu, Jianmin Lu, Sheng Yang
The association between obesity and reactive oxygen species (ROS) production in osteoarthritis (OA) patients has already been identified. However, the specific mechanism underlying ROS production and OA progression has never been elucidated. Osteoarthritic cartilage was obtained from patients undergoing total hip arthroplasty, and chondrocytes were isolated from these tissues. The cells were treated with varying concentrations (10,100,500 μM, and 5 mM) of oleic/palmitate (O/P) mixture at different times, that is at 6, 24, and 48 h...
July 2016: Annals of Clinical and Laboratory Science
https://www.readbyqxmd.com/read/27465875/mesenchymal-stem-cells-derived-from-human-bone-marrow-and-adipose-tissue-maintain-their-immunosuppressive-properties-after-chondrogenic-differentiation-role-of-hla-g
#20
Wen-Jing Du, Loic Reppel, Léonore Leger, Chantal Schenowitz, Celine Huselstein, Danièle Bensoussan, Edgardo D Carosella, Zhong-Chao Han, Nathalie Rouas-Freiss
Mesenchymal stem cells (MSC) have emerged as alternative sources of stem cells for regenerative medicine because of their multipotency and strong immune-regulatory properties. Also, human leukocyte antigen G (HLA-G) is an important mediator of MSC-mediated immunomodulation. However, it is unclear whether MSC retain their immune-privileged potential after differentiation. As promising candidates for cartilage tissue engineering, the immunogenic and immunomodulatory properties of chondro-differentiated MSC (chondro-MSC) require in-depth exploration...
October 1, 2016: Stem Cells and Development
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