scholarly journals THE USE OF MILLIPORE FILTERS IN ULTRASTRUCTURAL STUDIES OF CELL CULTURES AND VIRUSES

1968 ◽  
Vol 36 (1) ◽  
pp. 231-243 ◽  
Author(s):  
Robert M. McCombs ◽  
Matilda Benyesh-Melnick ◽  
J. Pierre Brunschwig
Keyword(s):  
Tissue Culture ◽  
Plasma Membranes ◽  
Fibroblast Cells ◽  
Virus Particles ◽  
Ultrastructural Studies ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Satellite Viruses ◽  
Murine Leukemia ◽  
Millipore Filters

A technique is described for embedding tissue culture cells that have been adsorbed or grown on Millipore filters. The acetone used during the embedding process rendered the filters transparent so that specific areas or cells could be chosen with the aid of the light microscope. Lymphoblastoid cells processed on the filters possessed well-defined plasma membranes and microvilli which were rarely present in cells from parallel cultures that were prepared by pelleting in the centrifuge. Fibroblast cells grown on filters retained their elongated appearance, in contrast to the rounded cells in pelleted preparations. Millipore filters were also used as a means of embedding virus pellets for sectioning. Preparations containing as few as 4 x 108 virus particles were suitable for study by the filter technique. Crude tissue-culture harvests of vaccinia virus and purified preparations of Rauscher murine leukemia and adeno-satellite viruses were successfully examined.

The Fine Structure of Replicating Simian Adenoviruses in LLC-MK2

1968 ◽  
Vol 26 ◽  
pp. 220-221
Author(s):  
Matias Pardo ◽  
Malcolm Slifkin ◽  
Leonard Merkow ◽  
Marie Sanchez
Keyword(s):  
Tissue Culture ◽  
Post Infection ◽  
Cesium Chloride ◽  
Longitudinal Section ◽  
Tubular Structures ◽  
Virus Particles ◽  
Ultrastructural Studies ◽  
Culture Cells ◽  
Simian Adenovirus ◽  
Infectivity Titer

The simian adenoviruses SV20, SV30 and SA7 have been found to be oncogenic in the Syrian hamster. The growth characteristics and replicative cycle of these viruses in tissue culture therefore appeared appropriate to investigate. Cesium chloride purified simian adenovirus with an infectivity titer of 100 TCID50, was inoculated into monolayers of LLC-MK2 cells. Cells were fixed in osmium tetroxide and embedded for ultrastructural studies at 1, 3, 6, 9, 18, 24, 48, 72, 120 and 192 hours post-infection.At the first hour post-infection, virus particles were adsorbed to the plasmalemma and found within the peripheral cytoplasm of many LLC-MK2 cells (Fig. 1). Although the first detection of infectious virus occurred at 14 hours and infectivity titers did not reach a maximum until 30 hours, intranuclear virus particles were observed by 3 hours in typical adenovirus crystalline array (Fig. 2) by means of electron microscopy. These typical honeycomb arrayed virus particles at 3 hours provided evidence of significant replication in approximately 5 percent of tissue culture cells examined. Simultaneously, a classical nuclear inclusion manifested by peripheral condensation of nuclear chromatin was evident by light microscopy. As early at 6 to 9 hours, unusual intranuclear concentric membranes formed “tubes” which contained linear arranged virus particles (Fig. 3). In transverse or tangential sections, these “tubes” appeared cochlear-like in shape. In longitudinal section, these intranuclear tubular structures contained individual virus particles at various stages of maturation in a linear arranged order. This arrangement resembled “peas in a pod”.

Labeling of Animal Cells with Fluorescent Dansyl Cerebroside

1976 ◽  
Vol 31 (11-12) ◽  
pp. 737-740b ◽  
Author(s):  
Richard T. C. Huang
Keyword(s):  
Tissue Culture ◽  
Plasma Membranes ◽  
Membrane Lipid ◽  
Animal Cells ◽  
Bhk Cells ◽  
Nuclear Membranes ◽  
Fluorescent Microscope ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Chicken Erythrocytes

Abstract A dansyl (diaminonaphthalenesulfonyl)-derivative of cerebroside was prepared which could be effectively incorporated into the plasma membranes of tissue culture cells and erythrocytes. The cells which had assimilated the glycolipid fluoresced intensely and could be observed under a fluorescent microscope. Cells were initially labeled rather homogeneously over the whole surface. With longer incubation time organization of the fluorescent glycolipid took place and patches of the lipid in the membrane were formed. The redistribution and organization of the membrane lipid could be demonstrated most clearly when cells labeled with this fluorescent glycolipid were infected with myxoviruses. After infection of MDBK and BHK cells with fowl plague virus areas of dense fluorescence appeared at margines of neighboring cells. When BHK cells were infected with Newcastle disease virus fusion of the cells was accompanied by complete redistribution of the glycolipid. Erythrocytes could also easily incorporate dansyl cerebroside. Chicken erythrocytes which contain cytoplasmic and nuclear membranes incorporated the fluorescent glycolipid in both membranes.

Transfer of Glycolipid between Membranes of Tissue Culture Cells, Using Dansylcerebroside as a Model

1977 ◽  
Vol 32 (5-6) ◽  
pp. 379-383 ◽  
Author(s):  
Richard T. C. Huang
Keyword(s):  
Tissue Culture ◽  
Plasma Membranes ◽  
Cellular Membranes ◽  
Bhk Cells ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Donor Cells ◽  
Mdbk Cells ◽  
Mode Of Attachment

Abstract Donor cells, which had incorporated dansylcerebroside in their membranes, could further transfer this glycolipid to monolayers of acceptor cells. The ease of transfer varied among acceptor cells, BHK cells being the best and MDBK cells the poorest acceptors of the cells tested. The process of transfer seemed to be mediated by lipids rather than by proteins of the membranes. The mode of attachment between donors and acceptors, such as classified as loose contact, tight adhesion or binding by lectin, did not significantly influence the extent of glycolipid transfer. However, modification of plasma membranes by infection of acceptor cells with myxoviruses re­sulted in enhancement of glycolipid transfer in some cases. Various factors have been evaluated with respect to dynamics of cellular membranes.

scholarly journals PEPSIN DIGESTION OF VIRUS PARTICLES IN CANINE HEPATITIS USING EPON-EMBEDDED MATERIAL

1967 ◽  
Vol 15 (11) ◽  
pp. 688-694 ◽  
Author(s):  
KATHLEEN F. GIVAN ◽  
CHARLOTTE TURNBULL ◽  
ANNE-MARIE JÉZÉQUEL
Keyword(s):  
Tissue Culture ◽  
Electron Microscope ◽  
Hepatitis Virus ◽  
Selective Extraction ◽  
Liver Cells ◽  
Pepsin Digestion ◽  
Virus Particles ◽  
Culture Cells ◽  
Tissue Culture Cells

Sixteen-week-old dogs were inoculated with 20,000 tissue culture doses (TCD50) of infectious canine hepatitis virus. Four days after inoculation, liver cells examined in the electron microscope contained both virus particles and nonviral paracrystalline formations. Tissue culture cells also contained virus particles and nonviral paracrystalline formations 48 hr after inoculation with virus. When Epon-embedded sections were treated for 1 hr with 0.5% pepsin in 0.1 N HCl, a selective extraction of the viral coats and of the nonviral paracrystals was observed.

Induction of Immunity in Mice with Tissue Culture Cells Chronically Infected with Rauscher Murine Leukemia Virus

1968 ◽  
Vol 128 (4) ◽  
pp. 1088-1092 ◽  
Author(s):  
S. A. Mayyasi ◽  
H. F. Foster ◽  
L. M. Bulfone ◽  
B. S. Wright ◽  
G. P. Shibley
Keyword(s):  
Tissue Culture ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Murine Leukemia

Tissue Culture Cells on Deformable Substrata: Biomechanical Implications

1984 ◽  
Vol 106 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Albert K. Harris
Keyword(s):  
Tissue Culture ◽  
Thin Layer ◽  
Silicone Rubber ◽  
Plasma Membranes ◽  
Time Lapse ◽  
The Body ◽  
Polydimethyl Siloxane ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Body Move

A method has been developed for the study of the forces which individual cells exert during their locomotion. Polydimethyl-siloxane (silicone fluid) was crosslinked on its surface by brief flaming to form a thin layer of silicone rubber. Tissue culture cells of many types were then plated out onto these rubber substrata and the propulsive forces these cells exert as they adhere and spread became visible as wrinkles and other distortions in the rubber. From time-lapse films of these distortions, it appears that the component cells of the body move by exerting shearing forces through their plasma membranes. How these forces are exerted and how this technique for observing them could be made more quantitative are discussed.

Recent Studies on a Murine Leukemia Virus Grown in Tissue Culture

1967 ◽  
Vol 25 ◽  
pp. 106-107
Author(s):  
L. Z. de Tkaczevski ◽  
E. de Harven ◽  
C. Friend
Keyword(s):  
Tissue Culture ◽  
Solid Tumors ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Supernatant Fluid ◽  
Virus Particles ◽  
Friend Leukemia ◽  
Type C ◽  
Leukemia Viruses ◽  
Murine Leukemia

Despite extensive studies, the correlation between the morphology and pathogenicity of murine leukemia viruses (MLV) has not yet been clarified. The virus particles found in the plasma of leukemic mice belong to 2 distinct groups, 1 or 2% of them being enveloped A particles and the vast majority being of type C. It is generally believed that these 2 types of particles represent different phases in the development of the same virus. Particles of type A have been thought to be an earlier form of type C particles. One of the tissue culture lines established from Friend leukemia solid tumors has provided the material for the present study. The supernatant fluid of the line designated C-1A contains an almost pure population of A particles as illustrated in Figure 1. The ratio is, therefore, the reverse of what is unvariably observed in the plasma of leukemic mice where C particles predominate.

Cytoplasmic Tubules in Cells Infected with Laryngotracheitis Virus

1969 ◽  
Vol 27 ◽  
pp. 376-377
Author(s):  
A. M. Watrach
Keyword(s):  
Tissue Culture ◽  
Small Proportion ◽  
Chicken Embryo ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Morphologic Characteristics ◽  
Infectious Laryngotracheitis ◽  
Laryngotracheitis Virus ◽  
In Cells

During a study of the development of infectious laryngotracheitis (LT) virus in tissue culture cells, unusual tubular formations were found in the cytoplasm of a small proportion of the affected cells. It is the purpose of this report to describe the morphologic characteristics of the tubules and to discuss their possible association with the development of virus.The source and maintenance of the strain of LT virus have been described. Prior to this study, the virus was passed several times in chicken embryo kidney (CEK) tissue culture cells.

Sign in / Sign up

Export Citation Format

Share Document