In Vitro Pulmonary Cell Culture in Pharmaceutical Inhalation Aerosol Delivery: 2-D, 3-D, and In Situ Bioimpactor Models

2016 ◽  
Vol 22 (17) ◽  
pp. 2522-2531 ◽  
Author(s):  
Maria F. Acosta ◽  
Priya Muralidharan ◽  
Samantha A. Meenach ◽  
Don Hayes ◽  
Stephen M.- Black ◽  
...  
Keyword(s):  
Cell Culture ◽  
Aerosol Delivery

scholarly journals A portable cell culture system with a simple microscope, oxygen sensor and MEA amplifier

2019 ◽  
Author(s):  
Dhanesh Kattipparambil Rajan
Keyword(s):  
Cell Culture ◽  
Chemical Sensing ◽  
Cell Imaging ◽  
Modular System ◽  
Disease Models ◽  
Environment Control ◽  
In Situ Imaging

Cell culture in-vitro is a well-known method to develop cell and disease models for studying physiologically relevant mechanisms and responses for various applications in life sciences. Conventional methods for instance, using static culture flasks or well plates, have limitations, as these cannot provide accurate tractable models for advanced studies. However, microscale systems can overcome this since they mimic the cells' natural microenvironment adequately. We have developed a portable live-cell imaging system with an invert-upright-convertible architecture and a mini-bioreactor for long-term simultaneous cell imaging and analysis, chemical sensing and electrophysiological recording. Our system integrates biocompatible cell-friendly materials with modular measurement schemes and precise environment control and can be automated. High quality time-lapse cell imaging is hugely useful in cell/disease models. However, integration of advanced in-vitro systems into benchtop microscopes for in-situ imaging is tricky and challenging. This is especially true with device based biological systems, such as lab/organ/body-on-chips, or mini-bioreactors/microfluidic systems. They face issues ranging from optical and physical geometry incompatibilities to difficulties in connectivity of flow and perfusion systems. However, the novel modular system we have developed either as an inverted or as an upright system can easily accommodate virtually any in-vitro devices. Furthermore, it can accept additional sensor or measurement devices quite freely. Cell characterization, differentiation, chemical sensing, drug screening, microelectrode-array-electrophysiological recordings, and cell stimulation can be carried out with simultaneous in-situ imaging and analysis. Moreover, our system can be configured to capture images from regions that are otherwise inaccessible by conventional microscopes, for example, cells cultured on physical or biochemical sensor systems. We demonstrate the system for video-based beating analysis of cardiomyocytes, cell orientation analysis on nanocellulose, and simultaneous long-term in-situ microscopy with pO2 and temperature sensing. The compact microscope as such is comparable to standard phase-contrast-microscopes without any detectable aberrations and is useful practically for any in-situ microscopy demands.

Partner telomeres during anaphase in crane-fly spermatocytes are connected by an elastic tether that exerts a backward force and resists poleward motion

2002 ◽  
Vol 115 (7) ◽  
pp. 1541-1549 ◽  
Author(s):  
James R. LaFountain ◽  
Richard W. Cole ◽  
Conly L. Rieder
Keyword(s):  
Cell Culture ◽  
Strong Evidence ◽  
Laser Microbeam ◽  
In Vitro Cell Culture ◽  
Pulling Forces ◽  
As If

As chromosomes move polewards during anaphase in crane-fly spermatocytes,trailing arms commonly stretch backwards for a brief time, as if tethered to their partners. To test that notion, a laser microbeam was used to sever trailing arms and thereby release telomere-containing arm segments (called acentric fragments because they lack kinetochores) from segregating chromosomes. Analysis of the movement of acentric fragments after their release provided clear evidence that previously conjoined partners were indeed tethered at their telomeres and that tethers exerted backward forces that were sufficient to move the fragment across the equator and into the opposite half-spindle. To address concerns that tethers might be artifacts of in vitro cell culture, spermatocytes were fixed in situ, and stretched arms within fixed cells provided strong evidence for tethers in vivo. The substantial resistance that tethers impose on the poleward movement of chromosomes must normally be over-ridden by the poleward `pulling' forces exerted at kinetochores. In spermatocytes, poleward forces are supplied primarily by the`traction fibers' that are firmly attached to kinetochores through end-on attachments to the plus ends of kinetochore microtubules.

In Situ and In Vitro Identification and Characterization of Cardiac Ganglion Neurons in the Crab, Carcinus maenas

1999 ◽  
Vol 81 (6) ◽  
pp. 2964-2976 ◽  
Author(s):  
Michelle A. Saver ◽  
Jerrel L. Wilkens ◽  
Naweed I. Syed
Keyword(s):  
Cell Culture ◽  
Carcinus Maenas ◽  
Membrane Properties ◽  
Cardiac Ganglion ◽  
Shore Crabs ◽  
Ganglion Neurons ◽  
Identification And Characterization

In situ and in vitro identification and characterization of cardiac ganglion neurons in the crab, Carcinus maenas. The aim of this study was to investigate the intrinsic membrane properties and hormonal responses of individual central pattern generating neurons in the cardiac ganglion of the shore crab Carcinus maenas. Because the cardiac ganglion in this crustacean species is buried within the heart musculature and is therefore inaccessible for direct morphological and electrophysiological analysis, we developed two novel in vitro preparations. First, to make the ganglion accessible, we established a brief enzymatic treatment procedure that enabled us to isolate the entire cardiac ganglion, in the absence of muscle tissue. Second, a cell culture procedure was developed to isolate individual neurons in vitro. With the use of both isolated ganglionic and neuronal cell culture techniques, this study provides the first direct account of the neuroanatomy of the cardiac ganglion in shore crabs. We demonstrate that cultured neurons not only survived the isolation procedures, but that they also maintained their intrinsic membrane and transmitter response properties, similar to those seen in the intact ganglion. Specifically, we tested the peptides proctolin, crustacean cardioactive peptide, the FLRFamide-related peptide F2, and an amine (serotonin) on both isolated ganglion and in vitro culture neurons. We measured changes in neuronal burst rate, burst amplitude, pacemaker slope, and membrane potential oscillation amplitude in response to the above four hormones. Each hormone either increased neuronal activity in spontaneously bursting neurons, or induced a bursting pattern in quiescent cells. The in vitro cell culture system developed here now provides us with an excellent opportunity to elucidate cellular, synaptic and hormonal mechanisms by which cardiac activity is generated in shore crabs.

IGF-1 loaded injectable microspheres for potential repair of the infarcted myocardium

2020 ◽  
pp. 088532822094850
Author(s):  
Elisabetta Rosellini ◽  
Niccoletta Barbani ◽  
Caterina Frati ◽  
Denise Madeddu ◽  
Diana Massai ◽  
...  
Keyword(s):  
Cell Culture ◽  
Growth Factor ◽  
In Situ Polymerization ◽  
Chemical Imaging ◽  
Homogeneous Distribution ◽  
Insulin Like Growth Factor ◽  
Gelatin Microspheres

The use of injectable scaffolds to repair the infarcted heart is receiving great interest. Thermosensitive polymers, in situ polymerization, in situ cross-linking, and self-assembling peptides are the most investigated approaches to obtain injectability. Aim of the present work was the preparation and characterization of a novel bioactive scaffold, in form of injectable microspheres, for cardiac repair. Gellan/gelatin microspheres were prepared by a water-in-oil emulsion and loaded by adsorption with Insulin-like growth factor 1 to promote tissue regeneration. Obtained microspheres underwent morphological, physicochemical and biological characterization, including cell culture tests in static and dynamic conditions and in vivo tests. Morphological analysis of the microspheres showed a spherical shape, a microporous surface and an average diameter of 66 ± 17µm (under dry conditions) and 123 ± 24 µm (under wet conditions). Chemical Imaging analysis pointed out a homogeneous distribution of gellan, gelatin and Insulin-like growth factor-1 within the microsphere matrix. In vitro cell culture tests showed that the microspheres promoted rat cardiac progenitor cells adhesion, and cluster formation. After dynamic suspension culture within an impeller-free bioreactor, cells still adhered to microspheres, spreading their cytoplasm over microsphere surface. Intramyocardial administration of microspheres in a cryoinjury rat model attenuated chamber dilatation, myocardial damage and fibrosis and improved cell homing. Overall, the findings of this study confirm that the produced microspheres display morphological, physicochemical, functional and biological properties potentially adequate for future applications as injectable scaffold for cardiac tissue engineering.

scholarly journals Formulation of Ocular In Situ Gels with Lithuanian Royal Jelly and Their Biopharmaceutical Evaluation In Vitro

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3552
Author(s):  
Kristina Perminaite ◽  
Mindaugas Marksa ◽  
Monika Stančiauskaitė ◽  
Tadas Juknius ◽  
Aidas Grigonis ◽  
...  
Keyword(s):  
Cell Culture ◽  
Refractive Index ◽  
Cell Viability ◽  
Royal Jelly ◽  
Biological Activities ◽  
Pharmaceutical Formulations ◽  
Exposure Test ◽  
Cell Culture Line

Royal jelly is a natural substance produced by worker bees that possesses a variety of biological activities, including antioxidant, anti-inflammatory, antibacterial, and protective. Although fresh royal jelly is kept at low temperatures, to increase its stability, it needs to be incorporated into pharmaceutical formulations, such as in situ gels. The aim of this study was to formulate in situ ocular gels containing Lithuanian royal jelly for topical corneal use in order to increase the retention time of the formulation on the ocular surface and bioavailability. Gels were evaluated for physicochemical characteristics (pH, rheological properties, refractive index) and in vitro drug release measuring the amount of 10-hydroxy-2-decenoic acid (10-HDA). An ocular irritation test and cell viability tests were performed using the SIRC (Statens Seruminstitut Rabbit Cornea) cell culture line. Results indicated that all the in situ gels were within an acceptable pH and refractive index range close to corneal properties. Rheology studies have shown that the gelation temperature varies between 25 and 32 °C, depending on the amount of poloxamers. The release studies have shown that the release of 10-HDA from in situ gels is more sustained than royal jelly suspension. All gel formulations were non-irritant according to the short-time exposure test (STE) using the SIRC cell culture line, and long-term cell viability studies indicated that the formulations used in small concentrations did not induce cell death. Prepared in situ gels containing royal jelly have potential for ocular drug delivery, and they may improve the bioavailability, stability of royal jelly, and formation of non-irritant ocular formulations.

scholarly journals A Review of the Effects of Major Atmospheric Pollutants on Pollen Grains, Pollen Content, and Allergenicity

2015 ◽  
Vol 2015 ◽  
pp. 1-29 ◽  
Author(s):  
Hélène Sénéchal ◽  
Nicolas Visez ◽  
Denis Charpin ◽  
Youcef Shahali ◽  
Gabriel Peltre ◽  
...  
Keyword(s):  
Cell Culture ◽  
Biological Effects ◽  
Pollen Grains ◽  
Atmospheric Pollutants ◽  
Combined Effects ◽  
Healthy Human ◽  
Pollen Content ◽  
The Impact

This review summarizes the available data related to the effects of air pollution on pollen grains from different plant species. Several studies carried out either onin situharvested pollen or on pollen exposed in different places more or less polluted are presented and discussed. The different experimental procedures used to monitor the impact of pollution on pollen grains and on various produced external or internal subparticles are listed. Physicochemical and biological effects of artificial pollution (gaseous and particulate) on pollen from different plants, in different laboratory conditions, are considered. The effects of polluted pollen grains, subparticles, and derived aeroallergens in animal models, inin vitrocell culture, on healthy human and allergic patients are described. Combined effects of atmospheric pollutants and pollen grains-derived biological material on allergic population are specifically discussed. Within the notion of “polluen,” some methodological biases are underlined and research tracks in this field are proposed.

Cell in situ zymography: an in vitro cytotechnology for localization of enzyme activity in cell culture

2012 ◽  
Vol 48 (8) ◽  
pp. 463-468 ◽  
Author(s):  
Aastha Chhabra ◽  
Astha Jaiswal ◽  
Umang Malhotra ◽  
Shrey Kohli ◽  
Vibha Rani
Keyword(s):  
Cell Culture ◽  
Enzyme Activity ◽  
In Situ Zymography

scholarly journals A portable cell culture system with a simple microscope, oxygen sensor and MEA amplifier

2019 ◽  
Author(s):  
Dhanesh Kattipparambil Rajan
Keyword(s):  
Cell Culture ◽  
Chemical Sensing ◽  
Live Cell Imaging ◽  
Oxygen Sensor ◽  
Cell Imaging ◽  
Imaging System ◽  
Electrophysiological Recording

Cell culture in-vitro is a well-known method to develop cell and disease models for studying physiologically relevant mechanisms and responses for various applications in life sciences. Conventional methods for instance, using static culture flasks or well plates, have limitations, as these cannot provide accurate tractable models for advanced studies. However, microscale systems can overcome this since they mimic the cells' natural microenvironment adequately. We have developed a portable live-cell imaging system with an invert-upright-convertible architecture and a mini-bioreactor for long-term simultaneous cell imaging and analysis, chemical sensing and electrophysiological recording. Our system integrates biocompatible cell-friendly materials with modular measurement schemes and precise environment control and can be automated. High quality time-lapse cell imaging is hugely useful in cell/disease models. However, integration of advanced in-vitro systems into benchtop microscopes for in-situ imaging is tricky and challenging. This is especially true with device based biological systems, such as lab/organ/body-on-chips, or mini-bioreactors/microfluidic systems. They face issues ranging from optical and physical geometry incompatibilities to difficulties in connectivity of flow and perfusion systems. However, the novel modular system we have developed either as an inverted or as an upright system can easily accommodate virtually any in-vitro devices. Furthermore, it can accept additional sensor or measurement devices quite freely. Cell characterization, differentiation, chemical sensing, drug screening, microelectrode-array-electrophysiological recordings, and cell stimulation can be carried out with simultaneous in-situ imaging and analysis. Moreover, our system can be configured to capture images from regions that are otherwise inaccessible by conventional microscopes, for example, cells cultured on physical or biochemical sensor systems. We demonstrate the system for video-based beating analysis of cardiomyocytes, cell orientation analysis on nanocellulose, and simultaneous long-term in-situ microscopy with pO2 and temperature sensing. The compact microscope as such is comparable to standard phase-contrast-microscopes without any detectable aberrations and is useful practically for any in-situ microscopy demands. [1] D. K. Rajan et al., "A Portable Live-Cell Imaging System With an Invert-Upright-Convertible Architecture and a Mini-Bioreactorfor Long-Term Simultaneous Cell Imaging, Chemical Sensing, and Electrophysiological Recording," in IEEE Access, vol. 6,pp. 11063-11075, 2018. doi: 10.1109/ACCESS.2018.2804378. [2] Antti-Juhana Mäki et al., "A Portable Microscale Cell Culture System with Indirect Temperature Control"SLAS TECHNOLOGY: Translating Life Sciences Innovation 2018 23:6, 566-579. [3] Hannu Välimäki et al.,Fluorimetric oxygen sensor with an efficient optical read-out for in vitro cellmodels,Sensors and Actuators B: Chemical,Volume 249,2017,pp.738-746,ISSN 0925-4005,https://doi.org/10.1016/j.snb.2017.04.182.

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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