Glucose oxidation by adherent and mechanically detached cultured cells

1991 ◽  
Vol 69 (10-11) ◽  
pp. 728-730 ◽  
Author(s):  
B. D. Honda ◽  
N. T. Glanville
Keyword(s):  
Glucose Oxidation ◽  
Spatial Organization ◽  
Cultured Cells ◽  
Test Tube ◽  
Growth Surface ◽  
Culture Flask ◽  
Cellular Architecture ◽  
Process Formation ◽  
Metabolic Properties ◽  
Diffusional Surface

This study examined the effect of mechanical detachment from the growth surface on energy metabolism of cultured cells. Oxidation of [1-14C]glucose measured by production of 14CO2 by adherent neuroblastoma (123 ± 5 nmol/mg protein per minute), glioma (128 ± 10 nmol/mg protein per minute), and fibroblast (137 ± 5 nmol/mg protein per minute) cultures was similar. Removing cells from the culture flask by scraping reduced glucose oxidation by 62, 30, and 82% in neuroblastoma, glioma, and fibroblast cultures, respectively. Transferring cells from a culture flask to a test tube, to control for diffusional surface area, did not further reduce glucose oxidation. Detaching cells from the growth surface destroyed the extensive process formation and disrupted the normal spatial organization on the culture plate. These results indicate that it is essential to maintain these aspects of cellular architecture when evaluating metabolic properties of cultured cells.Key words: glucose oxidation, neuroblastoma, glioma, fibroblasts.

Retraction of Thrombin-Induced Fibrin by Rhabdomyosarcoma BA 1112 Cultured Cells

1975 ◽  
Author(s):  
M. B. Donati ◽  
E. Dolfini ◽  
A. Cavenaghi ◽  
L. Morasen ◽  
G. de Gaetano
Keyword(s):  
Cultured Cells ◽  
Human Fibroblasts ◽  
Calf Serum ◽  
Specific Property ◽  
Test Tube ◽  
Cervix Carcinoma ◽  
Normal Epithelium ◽  
Cellular Integrity ◽  
Chang Liver ◽  
Human Cervix

The capacity to induce fibrin retraction has been considered a specific property of platelets until recently, when Niewiarowski et al. (Proc. Soc. Exp, Biol. Med. 140, 1199, 1972) observed fibrin retraction induced by human fibroblasts. As a part of a larger study on the interactions of cultured cells with fibrin, we have investigated the ability of the following cell lines to retract fibrin: KB (human oral epidermoid carcinoma); HeLa (human cervix carcinoma); Chang Liver (human, normal epithelium; Chang Conjunctiva (human normal epithelium); NCTC clone 929 (L) (fibroblasts from C3H/AN mice) and BA 1112 (rhabdomyosarcoma developed on WAG/Rji inbred rats). Cells were cultured in Eagle’s MEM, in Hank’s blanced salt solution plus 10% calf serum, removed from the flasks by trypsin treatment and resuspended at a concentration of 2 × 106/ml in Tyrode-albumin solution, containing Ca++ and Mg++. Human citrated platelet-poor plasma was clotted in a test tube at 37° C by thrombin in the presence of either the cell suspensions or buffer. Only BA 1112 cells were able to retract fibrin; the presence of Ca++, cellular integrity and random distribution in the sample were required for this activity. BA 1112 cells were able to modify the structuration of thrombin-induced fibrin as indicated by the marked increase of the maximal amplitude of thrombelastogram. BA 1112-induced fibrin retraction was inhibited by PGE1 and by some pyrimido-pyrimidine derivatives, not by apsirin. No retraction occurred when reptilase instead of thrombin was used as the clotting agent, even if the cells were preincubated with ADP. These results suggest that BA 1112 cells have a susceptibility to thrombin similar to that of platelets; this hypothesis is interesting in view of the muscular origin of these cells.(Supported by Grant NIH-PHRB-10RO1 CA 12764–01.)

scholarly journals Simplified in situ preparation of cultured cell monolayers for electron microscopy.

1980 ◽  
Vol 28 (2) ◽  
pp. 178-180 ◽  
Author(s):  
K Kawamoto ◽  
A Hirano ◽  
F Herz
Keyword(s):  
Electron Microscopy ◽  
Cultured Cells ◽  
Growth Surface ◽  
Cultured Cell ◽  
Cell Monolayers ◽  
In Situ Preparation

The use of xylene for the easy separation of cultured cells embedded in situ from their plastic growth surface is described. This step simplifies the preparation of cell monolayers for electron microscopy.

scholarly journals In vivo engineered extracellular matrix scaffolds with instructive niches for oriented tissue regeneration

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Meifeng Zhu ◽  
Wen Li ◽  
Xianhao Dong ◽  
Xingyu Yuan ◽  
Adam C. Midgley ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Tissue Regeneration ◽  
Spatial Organization ◽  
Cultured Cells ◽  
Functional Integration ◽  
Host Cells ◽  
Hierarchical Porous ◽  
Parallel Microchannels

Abstract Implanted scaffolds with inductive niches can facilitate the recruitment and differentiation of host cells, thereby enhancing endogenous tissue regeneration. Extracellular matrix (ECM) scaffolds derived from cultured cells or natural tissues exhibit superior biocompatibility and trigger favourable immune responses. However, the lack of hierarchical porous structure fails to provide cells with guidance cues for directional migration and spatial organization, and consequently limit the morpho-functional integration for oriented tissues. Here, we engineer ECM scaffolds with parallel microchannels (ECM-C) by subcutaneous implantation of sacrificial templates, followed by template removal and decellularization. The advantages of such ECM-C scaffolds are evidenced by close regulation of in vitro cell activities, and enhanced cell infiltration and vascularization upon in vivo implantation. We demonstrate the versatility and flexibility of these scaffolds by regenerating vascularized and innervated neo-muscle, vascularized neo-nerve and pulsatile neo-artery with functional integration. This strategy has potential to yield inducible biomaterials with applications across tissue engineering and regenerative medicine.

Morphological Changes in Tissue-Cultured Cells Due to Variation in Growth Matrices

1988 ◽  
Vol 46 ◽  
pp. 252-253
Author(s):  
M. E. Hogan ◽  
D. H. DeGaetano ◽  
K. L. Klomparens
Keyword(s):  
Tissue Culture ◽  
Cultured Cells ◽  
Cell Attachment ◽  
Morphological Changes ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Cell Types ◽  
Growth Surface ◽  
Textured Surface ◽  
Tissue Culture Plastic

As the use of isolated cell cultures increases as an experimental model, there has been a proportional increase in the number of matrices used for cell growth support. These matrices vary in shape, texture and porosity, and as a result, affect the cells grown on them.The ultrastructural characteristics of several of these growth matrices were examined using two cell types chosen for their distinct growth habits. Chinese Hamster Ovary cells and Balb 3T3 Mouse Fibroblasts were grown on flat substrates (glass, tissue culture plastic, Millipore filters) as well as spherical (glass, tissue culture plastic, cross-linked dextran) matrices. Cells were plated maintaining egual densities and growth surface area. Once the majority of the cells reached confluency (approx. one week), the cell morphology on each matrix was examined using scanning electron microscopy and digital analysis of cell attachment area.Variation in cell shape was dramatic between matrices, being most noticeable between a textured surface (filter, dextran bead), and that of a smooth (glass) surface (Figs. 1 and 2). Even within smooth surfaces, some variation was observed.

Whole Mount Observation of Cultured Cells

1980 ◽  
Vol 38 ◽  
pp. 802-805 ◽  
Author(s):  
K. Hama
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Image Quality ◽  
High Voltage ◽  
Cultured Cells ◽  
Specimen Thickness ◽  
High Voltage Electron Microscopy ◽  
Cellular Architecture ◽  
Voltage Electron ◽  
Whole Mount

The cellular architecture of cultured cells has been investigated on critical-point dried whole mount preparations with the aid of stereo-high voltage electron microscopy2,4,5. In these preparations, the absence of an embedding material permits an stereoobservation at rather low accelerating voltage1,3. In the present study, whole mount preparations of cultured chick fibroblasts were examined in the electron microscope operated at 100 KV, 200 KV, 500 KV, 750 KV and 1,000 KV to investigate the voltage dependency of the usable specimen thickness and of the image quality at different specimen thickness.

Nuclear lamina maintains global spatial organization of chromatin in Drosophila cultured cells

2018 ◽  
Author(s):  
Sergey V. Ulianov ◽  
Semen A. Doronin ◽  
Ekaterina E. Khrameeva ◽  
Pavel I. Kos ◽  
Sergey S. Starikov ◽  
...  
Keyword(s):  
Spatial Organization ◽  
Cultured Cells ◽  
Nuclear Lamina

A laser spectrofluorimeter for studies of cultured cells on the growth surface

1979 ◽  
Vol 92 (1) ◽  
pp. 74-81 ◽  
Author(s):  
Sandra H. Gianturco ◽  
Kong-Yi Hong ◽  
Marion R. Steiner ◽  
O.David Taunton ◽  
Richard L. Jackson ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Growth Surface

A method for measuring the oxygen consumption of intact cell monolayers

2000 ◽  
Vol 278 (4) ◽  
pp. L858-L863 ◽  
Author(s):  
Kamel Mamchaoui ◽  
Georges Saumon
Keyword(s):  
Culture Medium ◽  
Cultured Cells ◽  
Intact Cell ◽  
Second Law ◽  
Type Ii Cells ◽  
Type Ii ◽  
Cell Monolayers ◽  
Chamber Method ◽  
Metabolic Properties ◽  
Isolated Rat

This report describes an open-air method for measuring the O2 consumption (Q˙o 2) of intact monolayers of cultured cells. This method is based on Fick's second law of diffusion. It requires only a micromanipulator and a miniature O2 electrode to measure the[Formula: see text] gradient in the culture medium in the well. It was compared with the conventional oxygraph chamber method. Both methods gave the same value forQ˙o 2 in freshly isolated rat type II cells: 166 ± 15.3 nmol ⋅ h−1 ⋅ 106cells−1 for the open-air method and 151 ± 11.6 nmol ⋅ h−1 ⋅ 106cells−1 for the oxygraph chamber method ( n = 11 experiments). But the open-air method gave significantly larger values for Q˙o 2 in cells cultured for 2 days (236 ± 8.8 nmol ⋅ h−1 ⋅ 106cells−1) than the oxygraph method (71 ± 15.2 nmol ⋅ h−1 ⋅ 106cells−1; P < 0.001; n = 12 experiments). This suggests that the way cells are detached from their substratum to be placed in the oxygraph chamber affects theirQ˙o 2. The open-air method may be useful for studies on the metabolic properties of monolayers because the cells do not risk being damaged.

Glycolytic and oxidative metabolism in primary renal proximal tubule cultures

1989 ◽  
Vol 257 (2) ◽  
pp. C333-C340 ◽  
Author(s):  
K. G. Dickman ◽  
L. J. Mandel
Keyword(s):  
Proximal Tubule ◽  
Oxidative Metabolism ◽  
Cultured Cells ◽  
Lactate Production ◽  
Culture Conditions ◽  
Renal Proximal Tubule ◽  
Glycolytic Activity ◽  
Metabolic Properties ◽  
Primary Rabbit

Cultured cells often exhibit alterations in energy metabolism (increased glycolytic activity and decreased oxidative metabolism) during adaptation to the culture environment. The role of hypoxia as a mediator of these effects was examined by comparison of metabolism in primary rabbit renal proximal tubule (RPT) cultures maintained in stationary culture dishes (DISH), shaking Erlenmeyer flasks (SHAKE), and DISH cultures transferred back to SHAKE conditions (RESHAKE). Both oxidative metabolism and transport capacity were fully preserved in SHAKE cultures over a 24-h period. In contrast, within 6 h, DISH cultures exhibited a continuous decline in transport-dependent and -independent oxygen consumption, respiratory capacity, and ATP and K+ contents. The loss of oxidative metabolism in DISH cultures was accompanied by stimulation of lactate production, detectable within 1 h after plating. Comparison of metabolic properties of DISH cultures to those of RPT exposed to graded levels of hypoxia suggested that medium oxygen tensions may be as low as 1-3% in DISH cultures. RESHAKE cultures exhibited metabolic properties comparable to those of SHAKE cultures, indicating reversibility of DISH culture metabolism on reoxygenation. We concluded that DISH cultures rapidly become hypoxic as a consequence of static culture conditions. Shaking suspension cultures may provide a more metabolically appropriate model for long-term in vitro studies.

scholarly journals Assaying three-dimensional cellular architecture using X-ray tomographic and correlated imaging approaches

2020 ◽  
Vol 295 (46) ◽  
pp. 15782-15793 ◽  
Author(s):  
Peter O. Bayguinov ◽  
Max R. Fisher ◽  
James A. J. Fitzpatrick
Keyword(s):  
Spatial Organization ◽  
Three Dimensional ◽  
Specific Protein ◽  
Multidimensional Data ◽  
Data Sets ◽  
X Rays ◽  
Correlative Microscopy ◽  
Protein Targets ◽  
X Ray ◽  
Cellular Architecture

Much of our understanding of the spatial organization of and interactions between cellular organelles and macromolecular complexes has been the result of imaging studies utilizing either light- or electron-based microscopic analyses. These classical approaches, while insightful, are nonetheless limited either by restrictions in resolution or by the sheer complexity of generating multidimensional data. Recent advances in the use and application of X-rays to acquire micro- and nanotomographic data sets offer an alternative methodology to visualize cellular architecture at the nanoscale. These new approaches allow for the subcellular analyses of unstained vitrified cells and three-dimensional localization of specific protein targets and have served as an essential tool in bridging light and electron correlative microscopy experiments. Here, we review the theory, instrumentation details, acquisition principles, and applications of both soft X-ray tomography and X-ray microscopy and how the use of these techniques offers a succinct means of analyzing three-dimensional cellular architecture. We discuss some of the recent work that has taken advantage of these approaches and detail how they have become integral in correlative microscopy workflows.

Sign in / Sign up

Export Citation Format

Share Document