A micro tissue culture test for the titration and neutralization of rubella virus

1969 ◽  
Vol 15 (1) ◽  
pp. 67-71 ◽  
Author(s):  
John Furesz ◽  
Pierre Moreau ◽  
Walter Yarosh
Keyword(s):  
Tissue Culture ◽  
Rubella Virus ◽  
Fetal Calf Serum ◽  
Constant Pressure ◽  
Calf Serum ◽  
Rabbit Kidney ◽  
Tissue Cultures ◽  
Virus Titration ◽  
One Step ◽  
Microneutralization Test

A simple and reproducible micro tissue culture assay has been devised in RK13 and LLC-RK1 rabbit kidney cells for the titration and neutralization of rubella virus. In this "one-step" assay all virus and serum dilutions were prepared with spiral loops in disposable microplates and tissue cultures suspended in medium 199 and 3% horse or fetal calf serum were added to the microcups simultaneously. Micro tissue cultures were kept in a humidified incubator (36 °C) under a constant pressure of 5% CO2 for 8 days and were read microscopically for viral cytopathic changes on the seventh and eighth day. The microneutralization test performed in LLC-RK1 cell cultures was shown to be a reliable method for the detection of small amounts of rubella antibodies in human sera.The micro assay may be also applied to the virus titration of live, attenuated rubella vaccines.

A RAPID MICRO TISSUE CULTURE ASSAY IN BS-C-1 CELLS FOR THE TITRATION AND NEUTRALIZATION OF MEASLES VIRUS

1967 ◽  
Vol 13 (3) ◽  
pp. 313-319 ◽  
Author(s):  
Pierre Moreau ◽  
John Furesz
Keyword(s):  
Tissue Culture ◽  
Measles Virus ◽  
Constant Pressure ◽  
Fetal Bovine Serum ◽  
Mineral Oil ◽  
Tissue Cultures ◽  
Kidney Cell Line ◽  
Virus Titration ◽  
Inverted Microscope ◽  
Fetal Bovine

A simple, rapid and reproducible micro tissue culture assay in the BS-C-1 (Cercopithecus monkey kidney) cell line was developed for the titration and neutralization of measles virus. Serial dilutions of virus (or serum) were made with spiral loops in disposable microplates and 8 × 103 BS-C-1 cells suspended in 0.05 ml of medium 199 with 2% fetal bovine serum were added to each dilution. After the addition of 0.1 ml of sterile mineral oil to each microculture, the plates were covered with a plexiglass sheet and placed in a humidified incubator (37 °C) under a constant pressure of 5% CO2 for S days. Micro tissue cultures were examined with the aid of an inverted microscope for viral cytopathic changes on the fourth and fifth day.The micro assay was applied to the virus titration of various live, attenuated measles vaccines, as well as to the antibody titration of human and animal sera following vaccination with either live or inactivated measles vaccines.

Turbomolecular Pumped Electron Microscopy of Rubella Virus (In BHK Celis)

1968 ◽  
Vol 26 ◽  
pp. 204-205
Author(s):  
A. B. Taylor ◽  
G. C. Cole ◽  
M. A. Holcomb ◽  
C. A. Baechler
Keyword(s):  
Electron Microscopy ◽  
Rubella Virus ◽  
Phosphate Buffer ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Basal Medium ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Input Multiplicity ◽  
Baby Hamster Kidney Cells

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

Culture of Human Laryngeal Papilloma Cells in Vitro

1982 ◽  
Vol 90 (6) ◽  
pp. 728-735 ◽  
Author(s):  
Bettie M. Steinberg ◽  
Allan L. Abramson ◽  
Raymond P. Meade
Keyword(s):  
Tissue Culture ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Collagen Gel ◽  
Primary Cells ◽  
Normal Epithelium ◽  
Normal Cells ◽  
Laryngeal Papilloma ◽  
Hydrated Collagen Gel

Human laryngeal epithelial cells have been grown in tissue culture in a hydrated collagen gel containing Nutrient Mixture F12 (Gibco) supplemented with 15% fetal calf serum and 10μg//mL hydrocortisone. Primary cells often remain viable in culture for more than six months. They can be serially transferred two to four times before senescence. Cells derived both from normal epithelium and from laryngeal papilloma have been successfully cultured. Papilloma cells appear to contain more perinuclear granules and form fewer tight junctions than normal cells.

Ultrastructure of 2-Bromergocryptine-Treated Human Prolactin-Secreting Microadenomas in Culture

1977 ◽  
Vol 35 ◽  
pp. 536-537
Author(s):  
D.E. Bredesen ◽  
D. Schomberg ◽  
R. Kramer ◽  
C. Hammond ◽  
K.S. McCarty
Keyword(s):  
Tissue Culture ◽  
Osmium Tetroxide ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
The Other ◽  
Serum Prolactin ◽  
Organ Cultures ◽  
Short Term ◽  
Human Prolactin ◽  
Transphenoidal Approach

Speculation concerning the mechanism by which 2-bromoergocryptine (CB 154) acts to decrease serum prolactin (PRL) levels has revolved around two basic hypotheses; one concerning the importance of primary hypothalamic effects; the other emphasizing direct hypophyseal effects. The potential for hypothalamic effects were reduced in these experiments by explanting PRL-secreting microadenomas in organ culture and short term monolayer tissue culture. Tissue was obtained immediately from the operating room from 6 patients operated on by the transphenoidal approach. Cultures were maintained in Ham's F12 Nutrient Medium, 90% by volume with 10% fetal calf serum. CB 154 was added at concentrations of 0, 0.5, or 5.0μg/ml. The organ cultures were maintained for 24 hours before the addition of CB 154 and then for 24 to 48 hours in the presence of inhibitor after which specimens were fixed at 4°C with 4% glutaraldehyde, 0.1M. Cacodylate, pH 7.2, and post-fixed with 1% osmium tetroxide.

Fish kidney cell line in response to heat shock

1992 ◽  
Vol 50 (1) ◽  
pp. 704-705
Author(s):  
Li-Chu Tung ◽  
Yung-Reui Chen ◽  
Shiu-Nan Chen ◽  
Guang-Hsiung Kuo
Keyword(s):  
Heat Shock ◽  
Cell Line ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Uranyl Acetate ◽  
Ultrastructural Changes ◽  
Heat Shock Treatment ◽  
Acanthopagrus Schlegeli ◽  
Cacodylate Buffer ◽  
Fish Kidney

In the present study, the ultrastructural changes of BPK cells, a fibroblast-like cell line, derived from the kidney of juvenile black porgy Acanthopagrus schlegeli, under heat shock treatment are described.The BPK cells were maintained in L-15 medium supplemented with 10% fetal calf serum and 0.15 M NaCl at 28|C2. The heating was carried out in precalibrated water baths. Monolayers of cells, grown on coverslips in parafilm-sealed petri dishes were submerged under water for 30 min at 40|C treatments. Cells were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer supplemented with 6.6% sucrose, postfixed in 1% OsO4 and flat embedded in Spurr’s resin. Silver section were cut parallel to the substratum, stained with uranyl acetate and Reynold’s lead citrate, and examined in a Hitachi H-600 electron microscope at 75 KV.

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