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Chondrocyte flow cytometry

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]
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 cells (HF-MSCs) proliferation whilemaintaining the multi-lineage 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...
October 5, 2016: Stem Cells and Development
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...
September 23, 2016: Molecular Medicine Reports
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...
2016: Cellular and Molecular Biology
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
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
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...
2016: Stem Cell Research & Therapy
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
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
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
Zhenzhen Yang, Gábor Vajta, Ying Xu, Jing Luan, Mufei Lin, Cong Liu, Jianing Tian, Hongwei Dou, Yong Li, Tianbin Liu, Yijie Zhang, Lin Li, Wenxian Yang, Lars Bolund, Huanming Yang, Yutao Du
Mesenchymal stem cells (MSCs) exhibited self-renewal and less differentiation, making the MSCs promising candidates for adult somatic cell nuclear transfer (SCNT). In this article, we tried to produce genome identical pigs through hand-made cloning (HMC), with MSCs and adult skin fibroblasts as donor cells. MSCs were derived from either adipose tissue or peripheral blood (aMSCs and bMSCs, respectively). MSCs usually showed the expression pattern of CD29, CD73, CD90, and CD105 together with lack of expression of the hematopoietic markers CD34and CD45...
August 2016: Cellular Reprogramming
Li Yuan, Hou-Qi Liu, Min-Juan Wu
Stem cells are used with increasing success in the treatment of renal tubular injury. However, whether mesenchymal stem cells (MSC) differentiate into renal tubular epithelial cells remains controversial. The aims of the present study were to observe the localization of human embryonic MSCs (hMSCs) in the kidneys of newborn mice, and to investigate hMSC differentiation into tubular epithelium. Primary culture hMSCs were derived from 4-7-week-old embryos and labeled with the cell membrane fluorescent dye PKH-26...
August 2016: Experimental and Therapeutic Medicine
Bi Liu, Ming Lei, Tao Hu, Fei Yu, De-Ming Xiao, Hao Kang
SRT1720, a new discovered drug, was reported to activate silent information regulator 1 (SIRT1) and inhibit the chondrocyte apoptosis. However, the underlying mechanism remains elusive. In the present study, the chondrocytes were extracted from the cartilage tissues of New Zealand white rabbits, cultured in the presence of sodium nitroprusside (SNP) (2.5 mmol/L) and divided into five groups: 1, 5, 10, and 20 μmol/L SRT1720 groups and blank control group (0 μmol/L SRT1720). MTT assay was used to detect the chondrocyte viability and proliferation, and DAPI staining and flow cytometry to measure the chondrocyte apoptosis...
June 2016: Journal of Huazhong University of Science and Technology. Medical Sciences
Nimantana He, Zhenguo Dong, Bing Zhu, Mingtu Nuo, Shorgan Bou, Dongjun Liu
In our previous work, we found that the Inner Mongolia Arbas Cashmere goat hair follicle stem cells (gHFSCs) can be successfully differentiated into adipocyte, chondrocyte, and osteocyte lineages. In this study, we further examined the expression of the pluripotency and stemness markers Oct4, Nanog, Sox2, AKP, and TERT in gHFSCs by immunocytochemistry, flow cytometry, real-time PCR, and Western blot. Immunofluorescent staining showed that the gHFSCs were positive for all five markers. Fluorescence-activated cell sorting (FACS) further analyzed the positive expression of Oct4, Nanog, and Sox2 in the gHFSCs...
August 2016: In Vitro Cellular & Developmental Biology. Animal
Lina Gölz, Ruth Andrea Simonis, Joana Reichelt, Helmut Stark, Matthias Frentzen, Jean-Pierre Allam, Rainer Probstmeier, Jochen Winter, Dominik Kraus
OBJECTIVES: Resin infiltrants have been successfully used in dental medicine preventing the progression of tooth decay in an early phase of caries development. ICON(®) is an infiltrant of low-viscosity which penetrates via dentinal tubules into the lesion in dependence of the demineralization depth. Hence, we performed an in vitro study to determine the effect of ICON(®) on human dental pulp stem cells (hDPSCs). METHODS: Using explant technique, primary hDPSCs were collected from extracted teeth...
August 2016: Dental Materials: Official Publication of the Academy of Dental Materials
Hao Huang, Ying-Yao Quan, Xiao-Ping Wang, Tong-Sheng Chen
Gold nanoparticles (AuNPs) have been widely used in biomedical science including antiarthritic agents, drug loading, and photothermal therapy. In this report, we studied the effects of AuNPs with diameters of 3, 13, and 45 nm, respectively, on rabbit articular chondrocytes. AuNPs were capped with citrate and their diameter and zeta potential were measured by dynamic light scattering (DLS). Cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay after the rabbit articular chondrocytes were pre-incubated with 3, 13, and 45 nm AuNPs, respectively, for 24 h...
December 2016: Nanoscale Research Letters
Caroline E Gargett, Shanti Gurung
No abstract text is available yet for this article.
June 2016: Biology of Reproduction
E Katsiani, A Garas, C Skentou, A Tsezou, C I Messini, K Dafopoulos, A Daponte, I E Messinis
Mesenchymal stem cells (MSCs) can be obtained from a variety of human tissues. MSCs derived from placental chorionic villi of the first trimester are likely to resemble, biologically, embryonic stem cells (ESC), due to the earlier development stage of placenta. In the present study long-term cultures of MSC-like cells were assessed in order to evaluate MSCs multipotent characteristics and molecular features during the period of culture. CV-cells obtained from 10 samples of chorionic villus displayed typical fibroblastoid morphology, undergone 20 passages during a period of 120 days, maintaining a stable karyotype throughout long term expansion...
September 2016: Cell and Tissue Banking
Elena Stocco, Silvia Barbon, Paolo Radossi, Senthilkumar Rajendran, Daniele Dalzoppo, Marina Bortolami, Andrea Bagno, Francesca Grandi, Pier Giorgio Gamba, Pier Paolo Parnigotto, Giuseppe Tagariello, Claudio Grandi
Haemophilic arthropathy is the major cause of disability in patients with haemophilia and, despite prophylaxis with coagulation factor concentrates, some patients still develop articular complications. We evaluate the feasibility of a tissue engineering approach to improve current clinical strategies for cartilage regeneration in haemophiliacs by using autologous chondrocytes (haemophilic chondrocytes; HaeCs). Little is known about articular chondrocytes from haemophilic patients and no characterisation has as yet been performed...
October 2016: Cell and Tissue Research
C Zhou, H Zheng, J A Buckwalter, J A Martin
OBJECTIVE: Articular cartilage harbors chondrogenic progenitor cells (CPCs), a population that responds chemotactically to cell death. Because this behavior is reminiscent of macrophages, we hypothesized that CPCs have macrophage-like capabilities for scavenging cell and tissue debris through phagocytosis. DESIGN: CPCs, chondrocytes, synoviocytes, and macrophages were cultured with fluorophore-labeled chondrocyte debris for 3, 6, 12, or 24 h. Debris internalization was quantified by confocal microscopy and flow cytometry...
September 2016: Osteoarthritis and Cartilage
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