The bovine alveolar macrophage. 1. Isolation, in vitro cultivation, ultrastructure, and phagocytosis

1973 ◽  
Vol 19 (10) ◽  
pp. 1207-1210 ◽  
Author(s):  
M. L. Fox
Keyword(s):  
Electron Microscopy ◽  
Staphylococcus Aureus ◽  
Alveolar Macrophage ◽  
Calf Serum ◽  
In Vitro Cultivation ◽  
Foetal Calf Serum ◽  
Isolated Cells ◽  
Large Numbers ◽  
Bovine Alveolar Macrophage

The bovine alveolar macrophage (BAM) was isolated in large numbers from lung washings and could be cultivated in vitro up to 63 days in medium 199, 20% foetal calf serum (FCS). Freshly isolated cells examined by electron microscopy (EM) were similar to those of the mouse. Cultured BAM prepared for EM were larger than fresh cells and contained discrete packets of homogenous material, possibly lipid. The BAM phagocytosed Staphylococcus aureus (FDA209P, type 42D) but the percentage of cells phagocytosing was reduced when FCS was not present, when cells were infected for 30 min compared with 45 min and when 2 × 107 CFU/ml compared with 2 × 108 CFU/ml were used to infect the cells.

In vitro cultivation and developmental cycle in culture of a parasitic dinoflagellate (Hematodinium sp.) associated with mortality of the Norway lobster (Nephrops norvegicus) in British waters

Parasitology ◽  
1998 ◽  
Vol 116 (2) ◽  
pp. 115-130 ◽  
Author(s):  
P. L. APPLETON ◽  
K. VICKERMAN
Keyword(s):  
Calf Serum ◽  
Aquatic Organisms ◽  
Fresh Medium ◽  
Developmental Cycle ◽  
In Vitro Cultivation ◽  
Foetal Calf Serum ◽  
Nephrops Norvegicus ◽  
Norway Lobster ◽  
Characteristic Cycle

Dinoflagellates are common and often important parasites of aquatic organisms, but their developmental cycles are poorly known and have not been established in in vitro culture. The parasitic dinoflagellate (Hematodinium sp.) associated with mortality of the Norway lobster (Nephrops norvegicus) in British waters has been cultivated in vitro in 10% foetal calf serum in a balanced Nephrops saline. In culture the parasite undergoes a characteristic cycle of development. Circulating sporoblasts from the host's haemolymph in vitro generate 2 kinds of flagellated uninucleate dinospore, macrospores and microspores, either of which will, after 5 weeks in fresh medium, germinate to produce multinucleate unattached filamentous trophonts. These trophonts multiply by fragmentation and growth and may be serially subcultured in this form, at 2 week intervals, indefinitely. If not subcultured, the filamentous trophonts give rise to colonies of radiating filaments (‘gorgonlocks’) which subsequently attach to the substratum to form flattened web-like ‘arachnoid’ multi-nucleate trophonts. Arachnoid trophonts become arachnoid sporonts when they synthesize trichocysts and flagellar hairs and may give rise to secondary arachnoid sporonts or to dinospores which initiate a new cycle.

The differentiation of mouse gonads in vitro: a light and electron microscopical study

Development ◽  
1986 ◽  
Vol 97 (1) ◽  
pp. 189-199
Author(s):  
Sarah Mackay ◽  
Robert A. Smith
Keyword(s):  
Electron Microscopy ◽  
Culture Medium ◽  
Light And Electron Microscopy ◽  
Calf Serum ◽  
Microscopical Study ◽  
Electron Microscopical Study ◽  
The Other ◽  
Foetal Calf Serum ◽  
Electron Microscopical

Indifferent urogenital complexes were excised from mouse foetuses assessed by developmental criteria as day 10·5 or 11. After 4 or 6–7 days in culture, complexes were fixed and examined by light and electron microscopy. The effect of culturing sexed complexes in mixed sex groups was investigated. The effect of the presence or absence of foetal calf serum in the culture medium was considered. No evidence of inhibition of one sex by the other was found. Ovaries developed further in cultures than testes.

Addition of hypoxanthine to culture media allows in vitro cultivation of Babesia bovis and B. bigemina at reduced serum concentrations

Parasitology ◽  
2001 ◽  
Vol 123 (4) ◽  
pp. 357-363 ◽  
Author(s):  
L. NEVES ◽  
H. F. CROSS ◽  
L. LOUREIRO ◽  
A. AKÇA ◽  
M. HOMMEL ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Culture Media ◽  
Calf Serum ◽  
Standard Technique ◽  
Babesia Bovis ◽  
In Vitro Cultivation ◽  
Phase System ◽  
Foetal Calf Serum ◽  
Serum Replacement

The microaerophilous stationary phase system (MASP) was introduced in 1980 and successfully used as a standard technique for Babesia bovis and B. bigemina in vitro culture. The percentage of serum in the medium and the dependence on specific serum donors have been recognized as important constraints both for immunochemical studies and for the logistics of culture routine. In the present study the supplementation of RPMI 1640 with hypoxanthine at a concentration between 50 and 200 μM has enabled patterns of growth of B. bovis and B. bigemina to be achieved comparable to the standard technique with a simultaneous reduction of serum concentration from 40% to 5%. With hypoxanthine-supplemented medium it was possible to either replace the bovine serum from a specific donor with horse serum or use commercial adult bovine serum or foetal calf serum at 10%. When the serum replacement media Albumax II and GF21 were used, the growth of both B. bovis and B. bigemina markedly decreased after 3×72 h cycles. However, when these species were cultivated in culture flasks previously coated with cells from a murine peritoneal lavage, continuous parasite growth was achieved.

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

1997 ◽  
Vol 10 (01) ◽  
pp. 6-11 ◽  
Author(s):  
R. F. Rosenbusch ◽  
L. C. Booth ◽  
L. A. Dahlgren
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Tendon Healing ◽  
Matrix Proteins ◽  
Isolated Cells ◽  
Superficial Digital Flexor Tendon ◽  
Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

Drug activity against Onchocerca gutturosa males in vitro: a model for chemotherapeutic research on onchocerciasis

1987 ◽  
Vol 61 (4) ◽  
pp. 271-281 ◽  
Author(s):  
Simon Townson ◽  
C. Connelly ◽  
A. Dobinson ◽  
R. Muller
Keyword(s):  
Culture System ◽  
Serum Proteins ◽  
Good Condition ◽  
Calf Serum ◽  
New Drugs ◽  
Foetal Calf Serum ◽  
Partial Recovery ◽  
Feeder Cells ◽  
Compound Identification

ABSTRACTAn in vitro system for chemotherapeutic research using adult male Onchocerca gutturosa has been developed as a model for O. volvulus. Using a culture system consisting of medium MEM+10% heat inactivated foetal calf serum (IFCS)+LLCMK2 (monkey kidney) feeder cells in an atmosphere of 5% CO2 in air, we examined the effects of a range of antiparasitic drugs on worm motility. Ivermectin, levamisole, furapyrimidone, Mel W, chloroquine, metrifonate, flubendazole, amoscanate and the Ciba-Geigy compounds CGP 6140, CGP 20′376 and CGI 17658 either immobilized or significantly reduced motility levels at a concentration of 5x10−5M or less within a 7-day period. Worms were affected at very low concentrations by ivermectin (effective conc. to reduce motility levels to 50% of controls, 3.14x10−8M), levamisole (7.95x10−8M), CGP 6140 (8.87x10−9M) and CGP 20′376 (2.78x10−8M). Difficulties were experienced in accurately repeating the immotile endpoint for levamisole due to an inconsistent partial recovery of motility. Over a 7-day period diethylcarbamazine had little effect on motility levels, while suramin caused a slight increase in activity compared to controls at some timepoints. Subsequent experiments demonstrated some differences in drug efficacy depending on the presence or absence of serum and feeder cells in the culture system probably because of drug avidly binding to serum proteins. However, serum and cells were found to be essential ingredients of the culture system to maintain worms in good condition, indicating that new drugs should be evaluated both in the presence and absence of serum and cells. Comparisons were made between the responses of O. gutturosa and Brugia pahangi to certain drugs and these species were found to significantly differ in their sensitivities to ivermectin and a novel compound (Wellcome), indicating that Onchocerca parasites should be used wherever possible for compound identification and development intended for the treatment of onchocerciasis. The in vitro system described here, using male O. gutturosa, provides a basis for further research and a practical alternative to O. volvulus.

scholarly journals ELECTRON MICROSCOPY OF PLASMA-CELL TUMORS OF THE MOUSE

1961 ◽  
Vol 9 (2) ◽  
pp. 369-381 ◽  
Author(s):  
D. F. Parsons ◽  
M. A. Bender ◽  
E. B. Darden ◽  
Guthrie T. Pratt ◽  
D. L. Lindsley
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Complex Structure ◽  
Granular Body ◽  
Virus Particles ◽  
Culture Cells ◽  
Large Numbers ◽  
Tumor Agent

The X5563 tumor has been grown in tissue culture. Cells similar to those of the original tumor migrated from the explant and attached to the glass walls of the culture vessels. Electron microscopy showed that large numbers of particles, similar in morphology to virus particles, were associated with these cells after 7 days of culture. The two principal types of particles found in the tumor in vivo appear to be present in vitro. Many more of these particles, however, were larger and showed a more complex structure. Whereas the particles were mainly localized inside endoplasmic reticulum or the Golgi zone in the tumors in vivo, in the tissue culture the majority of the particles were associated with the plasma membrane and were found outside of the cells. The relation of the particles to the granular body is discussed as well as a possible relation to the mammary tumor agent.

Maturation of porcine oocytes for 33 or 44 hours in vitro in the presence or absence of serum

1998 ◽  
Vol 1998 ◽  
pp. 180-180
Author(s):  
K. Rust ◽  
M.E. Staines ◽  
GJ. McCallum ◽  
N.S. Prathalingam ◽  
S.A. Edwards ◽  
...  
Keyword(s):  
Disease Transmission ◽  
Calf Serum ◽  
International Markets ◽  
Foetal Calf Serum ◽  
Maturation Time ◽  
Nuclear Maturation ◽  
Defined Media ◽  
Porcine Embryo ◽  
Disease Risks

Porcine embryo production in vitrois providing the impetus for the development of cryopreservation strategies aimed at welfare-friendly domestic and international marketing and movement of stock in a manner that minimises risks of disease transmission. In the context of disease risks, defined media, which avoid the use of serum and other biohazardous products, are likely to become essential in the production of embryos for international markets. In preparing for this situation, the present comparative study investigated in vitronuclear maturation of porcine oocytes in the presence of either foetal calf serum (FCS) or polyvinyl alcohol (PVA). In addition, the effect of restricting the maturation time to 33 rather than 44 hours was examined.

Oesophagostomum radiatum: inhibition of in vitro development by newborn calf serum

1990 ◽  
Vol 64 (1) ◽  
pp. 9-14
Author(s):  
I. J. East ◽  
C. J. Fitzgerald
Keyword(s):  
Inhibitory Action ◽  
Natural Infection ◽  
Calf Serum ◽  
Foetal Calf Serum ◽  
Newborn Calf Serum ◽  
In Vitro Development ◽  
Specific Antibodies ◽  
Serum Inhibition ◽  
Vitro Development

ABSTRACTOesophagostomum radiatum developed to fourth stage larvae after 14 days in in vitro culture. However, development was totally inhibited if the standard 50% foetal calf serum in the medium was replaced by newborn calf serum. Inhibition did not occur with serum from cattle immune to O. radiatum through natural infection or experimental vaccination irrespective of the titre of specific antibodies to O. radiatum in each serum. The inhibitory action of NCS could be abolished by heat treatment at 56°C for 1 h but not by dialysis or repeated freeze-thawing. The inhibition was not consistent with observed differences in the activity of 19 enzymes in the various sera or the absence of various thiol-containing stimulants of worm development.

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