Objectives: The crosstalk between chondrocytes and adipose-derived stem cells (ADSCs) could regulate the secretion of multiple growth factors. However, it is not clear how the paracrine action in co-culture systems af...
Objectives: The crosstalk between chondrocytes and adipose-derived stem cells (ADSCs) could regulate the secretion of multiple growth factors. However, it is not clear how the paracrine action in co-culture systems affect cell migration. This study focused on the changes of cell migration of ADSCs and chondrocytes in co-culture conditions. Materials and methods: Primary ADSCs and chondrocytes were isolated from Sprague-Dawley rat. Transwell co-culture systems, inoculated with ADSCs and chondrocytes, were established in vitro. The morphology of the cells was observed 7 days post-seeding by inverted phase-contrast microscope. Additionally, the cytoskeleton changes were investigated by immunofluorescence staining. To detect the abundance of Vinculin, we used immunofluorescence and Western blotting. Additionally, the expression level of MMP-2, Hey1 and Hes1 was examined to determine the mechanisms of co-culture-induced cell migration changes. Results: The migration of ADSCs and chondrocytes in co-culture conditions significantly decreased compared with that in mono-culture groups, accompanied by the decrease of filopodia and the expression level of MMP-2. Conclusions: The overall study showed that the migration of ADSCs and chondrocytes differs significantly depending on culture conditions. Moreover, the Notch signalling pathway may be involved in this process. Accordingly, by studying changes in migration caused by co-culture, we obtained new insight into the crosstalk between ADSCs and chondrocytes.
We report on a facile method to detect the aggregation and co-aggregation of peptides by tryptophan fluorescence spectroscopy. Peptide aggregates (PAs) play a pivotal role in neurodegenerative diseases, such as Alzhei...
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We report on a facile method to detect the aggregation and co-aggregation of peptides by tryptophan fluorescence spectroscopy. Peptide aggregates (PAs) play a pivotal role in neurodegenerative diseases, such as Alzheimer's and Parkinson's. The detection of the formation of aggregates, especially in the early stage, will facilitate the diagnosis and treatment of the associated disease. In this study, by choosing a tryptophan-containing peptide of EP2, we investigated its fluorescence spectroscopic characteristics in the process of PAs. The results showed that the intensity of emission spectra was significantly enhanced with the formation of PAs within 48 h. In addition, by employing EP2 as a fluorescence probe, we found that EP2 was able to effectively monitor the aggregation of other peptides/proteins that are otherwise difficult to detect with conventional approach. Thus, these preliminary data provide a promising diagnostic tool to detect the formation of PAs.
ObjectivesPhysiological oxygen tension plays a critical role in homoeostatic maintenance and development of endochondral bone. Based on the proximity between uncalcified cartilage and subchondral bone, and microchanne...
ObjectivesPhysiological oxygen tension plays a critical role in homoeostatic maintenance and development of endochondral bone. Based on the proximity between uncalcified cartilage and subchondral bone, and microchannels that serve as a message delivery network between them, we aimed to explore the influence of low oxygen tension on soluble factor secretion in both chondrocytes and osteoblasts, after co-culture. Materials and methodsContact co-culture was achieved for morphological observation using red fluorescent protein (RFP)-labelled chondrocytes and green fluorescent protein (GFP)-labelled osteoblasts, and non-contact co-culture achieved by transwell chambers. This was used to screen genetic variation of growth factors in hypoxia, including respective phenotypic markers, factors involving hypoxia and angiogenesis relationships, bone morphogenetic family proteins, and other general factors. ResultsWe observed a significant increase in chondrocyte size following co-culture, in both normoxia and hypoxia, but not of osteoblasts. Expression of Aggrecan in chondrocytes and alkaline phosphatase in osteoblasts was down-regulated under hypoxia following co-culture. Under hypoxia, we found that expression of hypoxia-inducible factor-1, vascular endothelial growth factor-A/B, VE-cadherin, bone morphogenetic protein-2, and insulin-like growth factor-1 in chondrocytes, increased, but HIF-1, platelet-derived growth factor, BMP-5/-6 and fibroblast growth factor-1 in osteoblasts, decreased. ConclusionsThese results not only indicate the importance of crosstalk between chondrocytes and osteoblasts but also improve our understanding of the mechanisms underlying homoeostatic maintenance of endochondral bone.
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