17 research outputs found
Successful establishment of primary small airway cell cultures in human lung transplantation
No full textBackground: The study of small airway diseases such as post-transplant bronchiolitis obliterans syndrome (BOS) is hampered by the difficulty in assessing peripheral airway function either physiologically or directly. Our aims were to develop robust methods for sampling small airway epithelial cells (SAEC) and to establish submerged SAEC cultures for downstream experimentation
Where Do The Bronchioles Go? Epithelial-Mesenchymal Transition (EMT) Induction And Prevention In Epithelial Cells From Lung Transplant Patients
No full textAdditional file 1 of A modular 3D printed microfluidic system: a potential solution for continuous cell harvesting in large-scale bioprocessing
No full textAdditional file 1. The design criteria, methodology and simulation sections of the microfluidic system, methodology of cell culture and properties measurement, supplementary figures of the microfluidic system characterisation and MSCs characterisation post-harvesting
Rapid and Continuous Cryopreservation of Stem Cells with a 3D Micromixer
No full textCryopreservation is the final step of stem cell production before the cryostorage of the product. Conventional methods of adding cryoprotecting agents (CPA) into the cells can be manual or automated with robotic arms. However, challenging issues with these methods at industrial-scale production are the insufficient mixing of cells and CPA, leading to damage of cells, discontinuous feeding, the batch-to-batch difference in products, and, occasionally, cross-contamination. Therefore, the current study proposes an alternative way to overcome the abovementioned challenges; a highly efficient micromixer for low-cost, continuous, labour-free, and automated mixing of stem cells with CPA solutions. Our results show that our micromixer provides a more homogenous mixing of cells and CPA compared to the manual mixing method, while the cell properties, including surface markers, differentiation potential, proliferation, morphology, and therapeutic potential, are well preserved
Bronchial brushings for investigating airway inflammation and remodelling
No full textAsthma is the commonest medical cause for hospital admission for children in Australia, affects more than 300 million people worldwide, and is incurable, severe in large number and refractory to treatment in many. However, there have been no new significant treatments despite intense research and billions of dollars. The advancement in our understanding in this disease has been limited due to its heterogeneity, genetic complexity and has severely been hampered particularly in children by the difficulty in obtaining relevant target organ tissue. This review attempts to provide an overview of the currently used and recently developed/adapted techniques used to obtain lung tissue with specific reference to the airway epithelium. © 2011 The Authors Respirology © 2011 Asian Pacific Society of Respirology
Mussel inspired ZIF8 microcarriers: a new approach for large-scale production of stem cells
No full text© The Royal Society of Chemistry 2020. Metal-organic frameworks (MOFs) have high porosity, large surface area, and tunable functionality and have been widely used for drug loading. The aim of this study was to exploit unique features of zeolitic imidazolate framework-8 (ZIF8) to develop an innovative composite microcarrier (MC) for human mesenchymal stem cells (hMSCs) adhesion and proliferation. ZIF8 MCs were prepared by immobilization of polydopamine/polyethyleneimine (PDA/PEI) and ZIF8 on the surface of polystyrene beads. The chemical properties of MCs such as coating stability and homogeneity were characterized by different techniques such as ATR-FTIR, XRD, EDX, SEM, and contact angle. The prepared MCs were tested using human adipose-derived mesenchymal stem cells (hADSCs) in both static and dynamic conditions, and results were compared to a commercially available MC (Star-Plus), polydopamine coated MCs and amine-functionalized MCs as a control. Results show that PDA/PEI/ZIF8 coated MCs (in short: ZIF8 MCs) provides an excellent biocompatible environment for hADSCs adhesion and growth. In conclusion, ZIF8 MCs represent suitable and low-cost support for hADSCs culture and expansion, which can maximize cell yield and viability while preserving hADSCs multipotency. The present findings have revealed this strategy has the potential for chemical and topographical modification of MCs in tissue engineering applications
Regional Differences in Susceptibiity of Bronchial Epithelium to Mesenchymal Transition and Inhibition by the Macrolide Antibiotic Azithromycin
No full textObjective: Dysregulated repair following epithelial injury is a key forerunner of disease in many organs, and the acquisition of a mesenchymal phenotype by the injured epithelial cells (epithelial to mesenchymal transition, EMT) may serve as a source of fibrosis. The macrolide antibiotic azithromycin and the DNA synthesis inhibitor mycophenolate are in clinical use but their mechanism of action remains unknown in post-transplant bronchiolitis obliterans syndrome (BOS). Here we determined if regional variation in the EMT response to TGF beta 1 underlies the bronchiolocentric fibrosis leading to BOS and whether EMT could be inhibited by azithromycin or mycophenolate
The airway epithelium is a direct source of matrix degrading enzymes in bronchiolitis obliterans syndrome
No full textBACKGROUND: Long-term survival after lung transplantation is hindered by the development of bronchiolitis obliterans syndrome (BOS), and recent evidence suggests that dysregulated epithelial repair may underlie its development. Because matrix metalloproteinase (MMP) -2 and MMP-9 secretion is integral to repair, we hypothesized that airway epithelial cells from patients with BOS would over-express these matrix-degrading enzymes
Changes in activity of MMP-2 and -9 in primary human small airway epithelial cells (SAEC) and large airway epithelial cells (LAEC) stimulated with TGFβ1 <i>in vitro</i>.
No full text<p>Both SAEC and LAEC cell cultures were stimulated with TGFβ1 (50 ng/ml) for 96 h and activity of secreted matrix metalloproteinase (MMP) −2 and −9 in the culture supernatant was measured by gelatin zymography. (A) Initial zymograms suggested a marked increase in the activity of MMP-2 and MMP-9 after 96 h of stimulation. Densitometric quantification of zymograms showed a significant increase in the activity of (B) MMP-2 & (C) MMP-9 in the supernatant in both SAEC and LAEC. Furthermore, the increase in MMP-2 and -9 activity was significantly higher in SAEC compared to LAEC at 96 hrs. (p = 0.0027 for MMP-2 and p = 0.0004 for MMP-9, n = 23). Data presented as median ± IQR.</p
The role TGFβ receptor II (TGFBRII) and of Smad3 activation as potential mechanisms underlying the differential susceptibility of small airway epithelial cells (SAEC) and large airway epithelial cells (LAEC) to TGFβ1 induced EMT.
No full text<p>(A) To test whether the increased susceptibility of SAEC to EMT due to an increased expression of TGFBRII, <i>ex vivo</i> samples from small and large airway brushings were analyzed for gene expression of TGFBRII qRT-PCR. No significant difference in TGFβRII expression between SAEC and LAEC was observed (p = 0.56) (n = 6). (B) To investigate if increased susceptibility of SAEC to EMT was due to differences in the activation of the Smad-dependent pathway, the rate of Smad3 phosphorylation (normalized to β-actin) post TGFβ1 stimulation was measured by Western blot analysis on cell lysates from stimulated SAEC and LAEC cultures. There was also no significant difference in the rate of Smad3 phosphorylation following TGFβ1 stimulation (p = 0.21, n = 6). Data represented as median ± IQR.</p
