scholarly journals Engineering a dynamic model of the alveolar interface for the study of aerosol deposition

2020 ◽  
Author(s):  
Roberta Nossa ◽  
Joana Costa ◽  
Ludovica Cacopardo ◽  
Arti Ahluwalia
Keyword(s):  
Dynamic Model ◽  
Culture Media ◽  
Drug Efficacy ◽  
Aerosol Deposition ◽  
Porous Membrane ◽  
Breathing Motion ◽  
Micro Particles ◽  
Layer V ◽  
Epithelial Lung Cells ◽  
Air Liquid Interface

Nano and micro particles are widely used in industrial, household and medicinal applications. To understand the interaction between particles and epithelial cells, we developed a dynamic model of the alveolar interface. This system, named DALI (Dynamic Model for the ALveolar Interface), is a modular bioreactor composed of two chambers divided by a porous membrane where epithelial lung cells are seeded. The membrane is the support of the alveolar barrier that separates the two compartments of the alveolus: the air and blood side. The system integrates the following elements: i) Air/Liquid interface, thanks to the two chambers divided by the membrane: ii) Cell culture media flow, thanks to the presence of a peristaltic pump; iii) Lung breathing motion, thanks to an airflow that allows the stretching of the membrane; iv) Aerosol deposition system, to study the effects of drug efficacy or particle toxicity on the epithelial layer; v) Quartz Crystal Microbalance, to quantify the amount of aerosolized particles.

scholarly journals PHAIR – A biosensor for pH measurement in air-liquid interface

2020 ◽  
Author(s):  
Mohammadhossein Dabaghi ◽  
Neda Saraei ◽  
Gang Xu ◽  
Abiram Chandiramohan ◽  
Jonas Yeung ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Silver Chloride ◽  
Reference Electrode ◽  
Airway Epithelial Cells ◽  
Liquid Interface ◽  
Small Scale ◽  
Human Airway ◽  
Air Liquid Interface

1AbstractIn many biological systems, pH can be used as a parameter to understand and study cell dynamics. However, measuring pH in live cell culture is limited by the sensor ion specificity, proximity to the cell surface, and scalability. Commercially available pH sensors are difficult to integrate into a small-scale cell culture system due to their size and are not cost-effective for disposable use. We made PHAIR - a new pH sensor that uses a micro-wire format to measure pH in vitro human airway cell culture. Tungsten micro-wires were used as the working electrodes, and silver micro-wires with a silver/silver chloride coating were used as a pseudo reference electrode. pH sensitivity, in a wide and narrow range, and stability of these sensors were tested in common standard buffer solutions as well as in culture media of human airway epithelial cells grown at the air-liquid interface in a 24 well cell culture plate. When measuring the pH of cells grown under basal and challenging conditions using PHAIR, cell viability and cytokine responses were not affected. Our results confirm that micro-wires-based sensors have the capacity for miniaturization, and detection of diverse ions while maintaining sensitivity. This suggests the broad application of PHAIR in various biological experimental settings.

scholarly journals PHAIR: a biosensor for pH measurement in air–liquid interface cell culture

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mohammadhossein Dabaghi ◽  
Neda Saraei ◽  
Gang Xu ◽  
Abiram Chandiramohan ◽  
Jonas Yeung ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Silver Chloride ◽  
Reference Electrode ◽  
Airway Epithelial Cells ◽  
Liquid Interface ◽  
Small Scale ◽  
Human Airway ◽  
Air Liquid Interface

AbstractIn many biological systems, pH can be used as a parameter to understand and study cell dynamics. However, measuring pH in live cell culture is limited by the sensor ion specificity, proximity to the cell surface, and scalability. Commercially available pH sensors are difficult to integrate into a small-scale cell culture system due to their size and are not cost-effective for disposable use. We made PHAIR—a new pH sensor that uses a micro-wire format to measure pH in vitro human airway cell culture. Tungsten micro-wires were used as the working electrodes, and silver micro-wires with a silver/silver chloride coating were used as a pseudo reference electrode. pH sensitivity, in a wide and narrow range, and stability of these sensors were tested in common standard buffer solutions as well as in culture media of human airway epithelial cells grown at the air–liquid interface in a 24 well cell culture plate. When measuring the pH of cells grown under basal and challenge conditions using PHAIR, cell viability and cytokine responses were not affected. Our results confirm that micro-wire-based sensors have the capacity for miniaturization and detection of diverse ions while maintaining sensitivity. This suggests the broad application of PHAIR in various biological experimental settings.

scholarly journals PHAIR – A Biosensor for pH Measurement in Air-liquid Interface

2020 ◽  
Author(s):  
Mohammadhossein Dabaghi ◽  
Neda Saraei ◽  
Gang Xu ◽  
Abiram Chandiramohan ◽  
Jonas Yeung ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Silver Chloride ◽  
Reference Electrode ◽  
Airway Epithelial Cells ◽  
Liquid Interface ◽  
Small Scale ◽  
Human Airway ◽  
Air Liquid Interface

Abstract In many biological systems, pH can be used as a parameter to understand and study cell dynamics. However, measuring pH in live cell culture is limited by the sensor ion specificity, proximity to the cell surface, and scalability. Commercially available pH sensors are difficult to integrate into a small-scale cell culture system due to their size and are not cost-effective for disposable use. We made PHAIR - a new pH sensor that uses a micro-wire format to measure pH in vitro human airway cell culture. Tungsten micro-wires were used as the working electrodes, and silver micro-wires with a silver/silver chloride coating were used as a pseudo reference electrode. pH sensitivity, in a wide and narrow range, and stability of these sensors were tested in common standard buffer solutions as well as in culture media of human airway epithelial cells grown at the air-liquid interface in a 24 well cell culture plate. When measuring the pH of cells grown under basal and challenging conditions using PHAIR, cell viability and cytokine responses were not affected. Our results confirm that micro-wires-based sensors have the capacity for miniaturization, and detection of diverse ions while maintaining sensitivity. This suggests the broad application of PHAIR in various biological experimental settings.

Influence of fibroblasts on epidermization by keratinocytes cultured on synthetic porous membrane (insert) at the air-liquid interface

1994 ◽  
Vol 10 (5-6) ◽  
pp. 361-365 ◽  
Author(s):  
M. Robert ◽  
M. S. Noel-Hudson ◽  
J. Font ◽  
M. Aubery ◽  
J. Wepierre
Keyword(s):  
Porous Membrane ◽  
Liquid Interface ◽  
Air Liquid Interface

Abstract 409: Size-dependent Acute Cellular Responses of Unbound Free Fatty Acids

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Eun-Soo Lee ◽  
Mi Kyoung Lee ◽  
Se-hwa Kim
Keyword(s):  
Fatty Acids ◽  
Cell Death ◽  
Free Fatty Acids ◽  
Culture Media ◽  
Cellular Responses ◽  
Protective Role ◽  
Fatty Acid Binding ◽  
Size Dependent ◽  
Micro Particles

Introduction The level of free fatty acids (FFAs) in serum determines the pathophysiological condition. Most FFAs are carried in albumin-bound forms, but very small portion is remained unbound. Due to the methodological limitations, there is little known about the effect of unbound form. Objectives: This study aimed to understand the effect of local increase of unbound FFAs. Methods and Results: We synthesized the nano- and micro-particles of linoleate (LNP and LMP respectively) in unbound forms. The sizes of linoleate particles in distilled water, in serum-free, and in serum-added culture media were measured by dynamic light scattering. We confirmed that the size of LNPs in serum-added media were ~ 200 nm in diameter without aggregation. By the treatments of LNPs and LMPs in macrophage cell line (RAW 264.7), we studied size-dependent cellular responses including viability, lipid storage, and intracellular translocation. Increased lipid droplets in LNPs-treated cells were analyzed based on label-free coherent anti-Stokes Raman scattering imaging. We also found that LNPs (250 μM), not LMPs, activated the translocation toward nucleus of fatty acid binding protein 4 (FABP4), a carrier of intracellular FFA, by western blotting and immunocytochemistry. It was noteworthy that the nano-sized unbound FFA could induce post-transcriptional gene expression unlikely micro-sized FFAs. In the cells whose FABP4 function was inhibited by the chemical inhibitor, the cell death was significantly increased through apoptosis in short exposed time. This cell death response was mediated by endoplasmic reticulum stress suggesting the protective role of FABP4 on the imbalanced FFA metabolisms. Conclusions: We showed the size-dependent cellular responses by nano- and micro-sized unbound FFAs and the protective role of FABP4 on the FFA metabolisms. These findings provide the insights into acute cellular responses of unbound FFA.

High-pressure freezing of cells in suspension for low-voltage scanning electron microscopy (LVSEM)

1991 ◽  
Vol 49 ◽  
pp. 64-65
Author(s):  
Marek Malecki ◽  
James Pawley ◽  
Hans Ris
Keyword(s):  
Electron Microscopy ◽  
High Pressure ◽  
Low Voltage ◽  
Culture Media ◽  
Cell Structure ◽  
Freeze Substitution ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Cell Culture Media ◽  
Voltage Scanning

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

Kalinin: A new epithelial basement membrane adhesion molecule

1991 ◽  
Vol 49 ◽  
pp. 178-179
Author(s):  
Douglas R. Keene ◽  
Gregory P. Lunstrum ◽  
Patricia Rousselle ◽  
Robert E. Burgeson
Keyword(s):  
Basement Membrane ◽  
Culture Media ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Keratinocytes ◽  
Positive Staining ◽  
Human Amnion ◽  
Epithelial Basement Membrane ◽  
Type Vii Collagen ◽  
Keratinocyte Cell ◽  
Epithelial Basement

A mouse monoclonal antibody produced from collagenase digests of human amnion was used by LM and TEM to study the distribution and ultrastructural features of an antigen present in epithelial tissues and in cultured human keratinocytes, and by immunoaffinity chromatography to partially purify the antigen from keratinocyte cell culture media.By immunofluorescence microscopy, the antigen displays a tissue distribution similar to type VII collagen; positive staining of the epithelial basement membrane is seen in skin, oral mucosa, trachea, esophagus, cornea, amnion and lung. Images from rotary shadowed preparations isolated by affinity chromatography demonstrate a population of rod-like molecules 107 nm in length, having pronounced globular domains at each end. Polyacrylamide gel electrophoresis suggests that the size of this molecule is approximately 440kDa, and that it is composed of three nonidentical chains disulfide bonded together.

Sign in / Sign up

Export Citation Format

Share Document