scholarly journals A simple and reliable method for the localization in cell culture of single identifiable cells for ultrastructural analyses.

1984 ◽  
Vol 32 (4) ◽  
pp. 444-446 ◽  
Author(s):  
K Morrison-Graham ◽  
P H O'Lague
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Reliable Method ◽  
Single Cells ◽  
Ultrastructural Analysis ◽  
Culture Dish ◽  
Simple Method ◽  
Monolayer Cultures ◽  
A Cell ◽  
Individual Grid

A simple method for relocating single cells in monolayer cultures for subsequent morphological or ultrastructural analysis is reported. This consists of producing, on the culture dish surface, a nontoxic carbon grid that is preserved during processing for either transmission (TEM) or scanning (SEM) electron microscopy. For TEM studies these grids are readily transferred along with the cells into the embedding plastic, and thus individual grid squares containing a cell(s) of interest can be quickly located, remounted, and sectioned. These grids may be useful for ultrastructural analyses of single cells previously studied electrophysiologically or after microinjection of macromolecules.

scholarly journals Localized mechanical stimulation of single cells with engineered spatio-temporal profile

Lab on a Chip ◽  
2018 ◽  
Vol 18 (19) ◽  
pp. 2955-2965 ◽  
Author(s):  
M. Monticelli ◽  
D. S. Jokhun ◽  
D. Petti ◽  
G. V. Shivashankar ◽  
R. Bertacco
Keyword(s):  
Cell Culture ◽  
Mechanical Stimulation ◽  
Single Cells ◽  
Mechanical Stimuli ◽  
Temporal Profile ◽  
A Cell ◽  
Spatio Temporal ◽  
Stimulation Of

We introduce a new platform for mechanobiology based on active substrates, made of Fe-coated polymeric micropillars, capable to apply mechanical stimuli with tunable spatio-temporal profile on a cell culture.

scholarly journals Bioelectronics-on-a-chip for cardio myoblast proliferation enhancement using electric field stimulation

2020 ◽  
Vol 24 (1) ◽  
Author(s):  
Ángel Aragón ◽  
María Cebro-Márquez ◽  
Eliseo Perez ◽  
Antonio Pazos ◽  
Ricardo Lage ◽  
...  
Keyword(s):  
Cell Culture ◽  
Electric Field ◽  
Cell Density ◽  
Electric Circuit ◽  
Field Stimulation ◽  
Culture Chamber ◽  
Culture Dish ◽  
Electric Field Stimulation ◽  
Myoblast Proliferation ◽  
A Cell

Abstract Background Cardio myoblast generation from conventional approaches is laborious and time-consuming. We present a bioelectronics on-a-chip for stimulating cells cardio myoblast proliferation during culture. Method The bioelectronics chip fabrication methodology involves two different process. In the first step, an aluminum layer of 200 nm is deposited over a soda-lime glass substrate using physical vapor deposition and selectively removed using a Q-switched Nd:YVO4 laser to create the electric tracks. To perform the experiments, we developed a biochip composed of a cell culture chamber fabricated with polydimethylsiloxane (PDMS) with a glass coverslip or a cell culture dish placed over the electric circuit tracks. By using such a glass cover slip or cell culture dish we avoid any toxic reactions caused by electrodes in the culture or may be degraded by electrochemical reactions with the cell medium, which is crucial to determine the effective cell-device coupling. Results The chip was used to study the effect of electric field stimulation of Rat ventricular cardiomyoblasts cells (H9c2). Results shows a remarkable increase in the number of H9c2 cells for the stimulated samples, where after 72 h the cell density double the cell density of control samples. Conclusions Cell proliferation of Rat ventricular cardiomyoblasts cells (H9c2) using the bioelectronics-on-a-chip was enhanced upon the electrical stimulation. The dependence on the geometrical characteristics of the electric circuit on the peak value and homogeneity of the electric field generated are analyzed and proper parameters to ensure a homogeneous electric field at the cell culture chamber are obtained. It can also be observed a high dependence of the electric field on the geometry of the electrostimulator circuit tracks and envisage the potential applications on electrophysiology studies, monitoring and modulate cellular behavior through the application of electric fields.

scholarly journals Formation of Large Scaffold-Free 3-D Aggregates in a Cell Culture Dish by Ultrasound Standing Wave Trapping

2019 ◽  
Vol 45 (5) ◽  
pp. 1306-1315 ◽  
Author(s):  
Misa Nakao ◽  
Chikahiro Imashiro ◽  
Taiki Kuribara ◽  
Yuta Kurashina ◽  
Kiichiro Totani ◽  
...  
Keyword(s):  
Cell Culture ◽  
Standing Wave ◽  
Culture Dish ◽  
Wave Trapping ◽  
A Cell ◽  
Large Scaffold

Effect of Cooling Stimulus on Collection Efficiency of Calf Chondrocytes Cultivated on Metal Surface

2017 ◽  
Vol 11 (6) ◽  
pp. 925-931 ◽  
Author(s):  
Yuta Kurashina ◽  
◽  
Shogo Miyata ◽  
Jun Komotori
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Culture ◽  
Cell Proliferation ◽  
Collection Efficiency ◽  
Trypsin Treatment ◽  
Culture Substrate ◽  
A Cell ◽  
Number Of Cells ◽  
Scanning Electron

A cell culture module capable of cooling stimulus to collect cells efficiently on a metal culture substrate was developed. We evaluated the cell collection ratio and morphology of the collected cells. Following a cooling stimulus (0°C) for 20 min, the number of collected cells was increased by 50% compared to that collected after trypsin treatment without pipetting from the metal culture substrate. Following the cooling stimulus, cells were observed by fluorescence microscopy and scanning electron microscopy; the cell filopodia were shrunken compared to non-cooling-stimulated cells. Furthermore, the combination of collagenase and cooling stimulation resulted in the collection of a comparable number of cells as that obtained using only trypsin. Thus, cell proliferation was improved compared to that following trypsin treatment. Therefore, this method can be applied for culturing cells that are susceptible to trypsin damage.

scholarly journals Ultrastructural Analysis of DNA Complexes During Transfection and Intracellular Transport

2003 ◽  
Vol 51 (9) ◽  
pp. 1237-1240 ◽  
Author(s):  
Régis Cartier ◽  
Maria Velinova ◽  
Cathleen Lehman ◽  
Bettina Erdmann ◽  
Regina Reszka
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Intracellular Transport ◽  
Ultrastructural Analysis ◽  
Cell Entry ◽  
Dna Complexes ◽  
Simple Method ◽  
Transmission Electron ◽  
Internalization Process ◽  
Entry Mechanism

We present a simple method based on transmission electron microscopy that allows investigation of the early steps of polyplex-mediated transfection without the use of labeled DNA. The ultrastructural analysis showed internalization of 0.2–1-μm aggregates composed of 30–50-nm subunits. In addition, new details of the internalization process were revealed, suggesting an unspecific cell entry mechanism of large DNA aggregates.

scholarly journals THU0058 ENHANCEMENT OF CARTILAGE REGENERATION EFFICIENCY WITH HUMAN ADIPOSE STEM CELL THREE-DIMENSIONAL SPHEROID

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 241.2-241
Author(s):  
J. Y. Ko ◽  
E. Lee ◽  
J. Kim ◽  
G. I. Im
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Single Cells ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Cartilage Regeneration ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Culture Dish

Background:3D (three-dimensional) cell culture technology has been researched steadily because of its high potential of biocompatibility compared to single cells since 1990s, and is being developed to 3D spheroids recently. Spheroids are considered to reflect the natural organization of cells better than 2D cell cultures, and stem cells spheroids have been studied extensively in therapeutic transplantation. Stem cells were considered as a method of replacing autologous chondrocyte in regenerative treatment of articular cartilage. Compared to conventional single cells, 3D cell culture is artificially created an environment similar to a living body in vitro so that all cells collectively, a cell culture model that allows growth or interaction with the environment. Therefore, the findings of this study indicate that enhancement of treatment efficiency of stem cells caused by potential of survival and proliferation of hASC spheroid in Osteoarthritis. In conclusion, spheroid positive subpopulation of hASCs has high cell proliferation and survival but not apoptosis and cell death potential, which may contribute to successful cartilage regeneration and the development of stem cell therapies in the future.Objectives:Studied for 3D spheroids to investigate the mechanism of enhancement of survival and proliferationof hASC (human adipose stem cells) spheroid, which may contribute to successful improvement of therapeutic efficacy of stem cells.Methods:Cell isolation and culture / 3D cell culture dish preparation / hASCs culture on 3D cell culture dish / Real-time PCR analysis / Western blotting / Alcian blue staining / ACLT + MM (Anterior cruciate ligament transection with Medial meniscectomy) model / In vivo fluorescence for cell tracking / In vivo effects of spheroids in OA joint / Histological analysis / Enzyme-linked immunosorbent assay (ELISA) results for inflamma -tory cytokines in rat synovial fluid / Statistical AnalysisResults:In order to see how the spheroid showed more residual than single, and how effective it was in actual cartilage regeneration, the result of paraffin tissues were confirmed by safranin O staining for each condition. The tendency of cartilage regeneration efficiency was good for spheroid. Although the differences between the single and spheroid groups were small, they reaffirmed that they could somewhat protect cartilage and help regeneration treatment. However, immunohistochemistry of HN(Human nucleic antigen) staining showed that cells of single and spheroid were not observed in the wound but disappeared by the paracrine effect.Conclusion:Spheroids do not exhibit differentiation characteristics, but they could be seen as a result of expression of related genes such as Bax, Bcl-XL and Alcian blue staining. Spheroids tend to have low potential of cell death rather than proliferation and reduction in the proliferation. So, we conclude the fact that instead of hASCs going directly to the surgical site to regenerate cartilage, they can help catrilage regeneration.Acknowledgments:This research was supported by the National Research Foundation of Korea (NRF-2019R1H1A2039685 and 2019R1I1A1A01043778).Disclosure of Interests:None declared

scholarly journals Bioelectronics-on-a-chip for cardio myoblast proliferation enhancement using electric field stimulation.

2020 ◽  
Author(s):  
Angel Aragon ◽  
María Cebro-Márquez ◽  
Eliseo Perez ◽  
Antonio Pazos ◽  
Ricardo Lage ◽  
...  
Keyword(s):  
Cell Culture ◽  
Electric Field ◽  
Cell Density ◽  
Electric Circuit ◽  
Field Stimulation ◽  
Culture Chamber ◽  
Culture Dish ◽  
Electric Field Stimulation ◽  
Myoblast Proliferation ◽  
A Cell

Abstract Background: Cardio myoblast generation from conventional approaches is laborious and time-consuming. We present a bioelectronics on-a-chip for stimulating cells cardio myoblast proliferation during culture. Method: The bioelectronics chip fabrication methodology involves two different process. In the first step, an aluminum layer of 200 nm is deposited over a soda-lime glass substrate using physical vapor deposition and selectively removed using a Q-switched Nd:YVO4 laser to create the electric tracks. To perform the experiments, we developed a biochip composed of a cell culture chamber fabricated with polydimethylsiloxane (PDMS) with a glass coverslip or a cell culture dish placed over the electric circuit tracks. By using such a glass cover slip or cell culture dish we avoid any toxic reactions caused by electrodes in the culture or may be degraded by electrochemical reactions with the cell medium, which is crucial to determine the effective cell-device coupling. Results: The chip was used to study the effect of electric field stimulation of Rat ventricular cardiomyoblasts cells (H9c2). Results shows a remarkable increase in the number of H9c2 cells for the stimulated samples, where after 72 hours the cell density double the cell density of control samples. Conclusions: Cell proliferation of Rat ventricular cardiomyoblasts cells (H9c2) using the bioelectronics-on-a-chip was enhanced upon the electrical stimulation. The dependence on the geometrical characteristics of the electric circuit on the peak value and homogeneity of the electric field generated are analyzed and proper parameters to ensure a homogeneous electric field at the cell culture chamber are obtained. It can also be observed a high dependence of the electric field on the geometry of the electrostimulator circuit tracks and envisage the potential applications on electrophysiology studies, monitoring and modulate cellular behavior through the application of electric fields

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