Laser-induced fusion of mammalian cells and plant protoplasts

1987 ◽  
Vol 88 (2) ◽  
pp. 145-149
Author(s):  
R. Wiegand ◽  
G. Weber ◽  
K. Zimmermann ◽  
S. Monajembashi ◽  
J. Wolfrum ◽  
...  
Keyword(s):  
Cell Culture ◽  
B Lymphocytes ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Laser Light ◽  
Plant Protoplasts ◽  
Myeloma Cells ◽  
Normal Culture ◽  
Laser Microbeam ◽  
Fusion Products

An ultraviolet-laser microbeam was shown to be suitable for inducing fusion of individually selected plant protoplasts or of B-lymphocytes with myeloma cells. The fusion took place in normal culture medium and the fusogenic condition perturbed the cells only for a fraction of a millisecond. Without manipulating the cell culture except for exposing the cells to laser light, fusion products between preselected individual pairs may be produced.

scholarly journals Adaptation of Cholesterol Requiring NS0 Cells to Serum Free Culture Conditions

2011 ◽  
Vol 12 (4) ◽  
Author(s):  
Junaid Muneer Raja ◽  
Nurina Anuar ◽  
And Badarulhisam Abdul Rahman
Keyword(s):  
Cell Culture ◽  
Monoclonal Antibodies ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Downstream Processing ◽  
Western World ◽  
Human Consumption ◽  
Cell Culture Medium ◽  
Serum Free ◽  
Ns0 Cells

Colorectal cancer is the third most common form of cancer and the second leading cause of cancer-related death in the Western world. The answers to such life threatening diseases and cancers are monoclonal antibodies (MAb's) which are widely used as therapeutic agents. World demand for currently approved MAb's is on the order of a few kilograms per year. However, new therapeutic MAb's are under development and require doses of several hundred milligrams to a gram over the course of therapy. Very often to cater for the special requirements for the growth of mammalian cells, serum is added to the cell culture medium. However, removal of serum from the cell culture medium is often carried out, especially if the end product is to be used for human consumption, in order to eliminate various disadvantages such as high physiological variability, high batch to batch variability, risk of contamination and high cost, and challenges posed in the downstream processing of the product. In this paper, the adaptation of cholesterol requiring NS0 cells to commercially available serum free media is presented. ABSTRAK: Kanser kolorektum merupakan kanser ketiga paling umum dan kini berada di tempat kedua penyebab kematian berkaitan kanser di negara Barat. Jawapan kepada penyakit yang mengancam nyawa dan penyakit kanser adalah antibodi monoklon (monoclonal antibodies ((MAb's)) yang digunakan sebagai agen terapeutik. Permintaan dunia terhadap MAb's yang diluluskan adalah dalam bilangan beberapa kilogram setahun. Namun, terapeutik MAb's yang baru adalah di bawah penyelidikan dan memerlukan beberapa ratus dos milligram hingga satu gram dalam satu peringkat terapi. Sering kali untuk memenuhi permintaan terhadap tumbesaran sel mamalia, serum dicampurkan dengan sel kultur perantara. Walaupun begitu, pemindahan serum dari sel kultur perantara sering dilakukan, terutamanya jika produk akhir digunakan untuk kegunaan manusia; untuk mengurangkan pelbagai kelemahan seperti kebolehubahan psikologi yang tinggi, kebolehubahan yang tinggi daripada satu kumpulan ke satu kumpulan lain, risiko pencemaran, kos yang tinggi, dan cabaran mendatang dalam pemprosesan produk. Dalam perbentangan ini, kolestrol yang diubah memerlukan sel NS0 yang dikomersilkan dengan serum bebas perantara.

GTSF-2: A new, versatile cell culture medium for diverse normal and transformed mammalian cells

1997 ◽  
Vol 33 (5) ◽  
pp. 344-351 ◽  
Author(s):  
Peter I. Lelkes ◽  
Esther Ramos ◽  
Victor V. Nikolaychik ◽  
Dawn M. Wankowski ◽  
Brian R. Unsworth ◽  
...  
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Cell Culture Medium

Serum effects on the response of mammalian cells to the exotoxins of Pseudomonas aeruginosa and Corynebacterium diphtheriae

1977 ◽  
Vol 23 (2) ◽  
pp. 175-182 ◽  
Author(s):  
John L. Middlebrook ◽  
Rebecca B. Dorland
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Transport Processes ◽  
Bacterial Toxins ◽  
Culture Studies ◽  
Probable Presence ◽  
Human Sera

The response of mammalian cells to Pseudomonas and diphtheria exotoxins was studied. A method was developed whereby the sensitivity of cells to these two toxins could be quantitated. The method is versatile and can be used to study the effects of toxins on many cellular metabolic or transport processes. The type of serum used in the culture medium significantly influenced the response of cells to the toxins. Calf, horse, and human sera protected cells while fetal calf serum did not. Precipitation with (NH4)2SO4 demonstrated the probable presence of toxin-specific antibody in the protective calf serum while none was detected in the nonprotective fetal calf serum. The level of antibody in calf serum, as titrated by hemagglutination, was sufficient to account for all the observed protection. It is suggested that fetal calf serum be used for all future cell culture studies of bacterial toxins.

The Use of Artemia Salina in Toxicity Testing

1992 ◽  
Vol 20 (2) ◽  
pp. 297-301 ◽  
Author(s):  
Lillemor Lewan ◽  
Marianne Andersson ◽  
Paloma Morales-Gomez
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Cultured Cells ◽  
Sea Water ◽  
Toxicity Testing ◽  
Artemia Salina ◽  
Good Survival ◽  
Cell Culture Medium ◽  
A Cell

This study shows that the Artemia assay, which is usually performed by incubation for a 24-hour period in artificial sea water, can also be performed in phosphate buffered saline (PBS) at pH 7.2, or in a cell culture medium, both of which are used in toxicity assays employing mammalian cells. Thus, by using the equivalent media for incubation, toxic effects in the Artemia assay and toxic mechanisms can more accurately be compared with results obtained in mammalian cell toxicity. Survival of control animals is good for 72 hours, provided that infection can be avoided. A pH greater than 6 is essential for good survival of Artemia Salina, and a pH greater than 10.5 should be avoided. Because of the risk of infection at low saline concentrations, a decreased incubation time of 16 or 12 hours is recommended. The lethal concentrations (LC10 and LC50) in the 24-hour Artemia assay of the first ten chemicals in the MEIC programme were measured in PBS, and the results compared with those from a previous study of the effects in a 10-minute acute ATP leakage assay, specifically indicating lesions in the cell membrane. The EC10 and EC50 values for the alcohols in the Artemia assay were 35–75% of the corresponding values in the ATP leakage assay. For paracetamol and amitriptyline, the EC10 and EC50 values in the Artemia assay were 2–16% of the corresponding values in the ATP-leakage assay. The greatly increased toxicity of the two compounds in the animals may be related to systemic effects. FeSO4 was very toxic to Artemia salina at a concentration of lug/ml, but it did not cause leakage of ATP from cultured cells, even at a concentration of 8,000μg/ml, showing that widely different mechanisms of interaction with FeSO4 are measured by the two assays. A lower toxicity of polygodial, a sesquiterpenoid unsaturated dialdehyde, in cell culture medium, was obvious in the Artemia assay. Thus, some factors in cell culture medium must, by interacting with the toxic molecule, protect the animals against the toxicity of polygodial, as was previously found in cultured cells.

scholarly journals Orchestrating Extracellular Vesicle With Dual Reporters for Imaging and Capturing in Mammalian Cell Culture

2021 ◽  
Vol 8 ◽  
Author(s):  
Daniel Levy ◽  
Mai Anh Do ◽  
Jiayi Zhang ◽  
Annie Brown ◽  
Biao Lu
Keyword(s):  
Cell Culture ◽  
Mammalian Cell ◽  
Cellular Uptake ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Magnetic Beads ◽  
Mammalian Cell Culture ◽  
Reporter System ◽  
Viral Glycoprotein ◽  
Dual Reporter

Background: Recent technological advancements have enabled live-cell imaging of intracellular organelles to monitor their biogenesis in mammalian cells. However, applying this method to gain insight into extracellular organelles, such as extracellular vesicles (EVs), presents unique challenges that require special considerations in design and engineering.Results: We have developed a dual-reporter system that combines genetic fusion, fluorescence microcopy and magnetic beads capture of EVs to study the biogenesis of EVs in mammalian cell cultures. First, we genetically produced a series of reporters by fusing a green fluorescent protein (GFP) and an affinity peptide (6xHis), with either the endogenous transmembrane protein, CD63, or EVs targeting vesicular stomatitis viral glycoprotein (VSVG). Transfection of these reporters into human 293T cells resulted in expression and integration of these reporters into pre-exosome compartments, which were subsequently released into the culture medium. Confocal imaging and nano-particle tracking analysis demonstrated that EVs were appropriately labeled and exhibited a single dominant peak in the 80–110 nm size range, indicating that isolated EVs were comprised of micro-vesicles and/or exosome subpopulations. Incubation of isolated EVs with nickel-coated magnetic beads resulted in successful capture of GFP-positive EVs. Finally, addition of EVs into culture medium was able to reveal the cellular uptake of GFP-labeled EVs by recipient cells. Taken together, our dual-reporter system provides a powerful method for both monitoring and capturing of EVs in mammalian cell culture systems.Conclusion: A dual-reporter system provides a robust tool to study the life cycle of EVs in mammalian cells from biogenesis and excretion to cellular uptake.

Examination of various cell culture techniques for co-incubation of virulent Treponema pallidum (Nichols I strain) under anaerobic conditions

1976 ◽  
Vol 4 (4) ◽  
pp. 360-371
Author(s):  
P L Sandok ◽  
S T Knight ◽  
H M Jenkin
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Treponema Pallidum ◽  
Dark Field ◽  
Anaerobic Conditions ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Dark Field Microscopy

Treponema pallidum (Nichols virulent) was incubated with and without cells in cell culture medium reduced to -275 mV Ecal, pH 7.3, under deoxygenated conditions. Five to ten percent of the treponemes attached to cells and remained motile for at least 120 h in cell-treponeme systems of co-incubation. Virulent treponemes could be detected after 120 to 144 h in the supernatant fluids of cell-treponeme co-incubation cultures and in cell-free tubes containing medium harvested from aerobically cultivated mammalian cells. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Increases in treponemal numbers were observed using dark-field microscopy but were not substantiated using the rabbit lesion test. Continuous passage of the treponeme was not achieved in vitro.

scholarly journals Long-Term Culture of Giardia lamblia in Cell Culture Medium Requires Intimate Association with Viable Mammalian Cells

2019 ◽  
Vol 87 (11) ◽  
Author(s):  
Theodore E. Nash
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Cell Lines ◽  
Giardia Lamblia ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Host Cells ◽  
Cell Culture Medium ◽  
Heterologous Cell

ABSTRACT Giardia lamblia is usually cultured axenically in TYI-S-33, a complex medium which does not permit survival and growth of mammalian cells. Likewise, medium commonly used to maintain and grow mammalian cells does not support healthy trophozoite survival for more than a few hours. The inability to coculture trophozoites and epithelial cells under optimal conditions limits studies of their interactions as well as interpretation of results. Trophozoites of the WB isolate but not the GS isolate were repeatedly adapted to grow stably in long-term cocultures with Caco2, Cos7, and mouse tumor rectal (RIT) cell lines using hybridoma-screened Dulbecco’s modified Eagle’s medium and 10% fetal calf serum. Giardia did not grow in spent cell culture medium or when separated by a permeable membrane using transwell methodology. Giardia chronically cocultured with specific cell lines became adapted (conditioned). These Giardia cocultures grew better than nonconditioned trophozoites, and the cell lines differed in their ability to support trophozoite growth in the order of RIT > Cos7 > Caco2. Trophozoites conditioned on one cell line and then grown in the presence of a heterologous cell line changed their growth rate to that seen in conditioned Giardia from the heterologous cell line. Trophozoite survival required intimate contact with cells, suggesting that trophozoites obtain an essential nutrient or growth factor from mammalian cells. This may explain why Giardia trophozoites adhere to the small intestinal epithelium during human and animal infections. This coculture system will be useful to understand the complex interactions between the host cells and parasite.

3-iodothyronamine (T1AM) is taken up and rapidly metabolized in cell culture medium only at low concentration

2020 ◽  
Author(s):  
Federica Saponaro ◽  
Marco Borsò ◽  
Sara Verlotta ◽  
Lavinia Bandini ◽  
Alessandro Saba ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Low Concentration

Effect of Atmospheric-Pressure Helium Plasma Jet on Cell Culture Medium

2013 ◽  
Vol 133 (5) ◽  
pp. 278-285
Author(s):  
Norimitsu Takamura ◽  
Douyan Wang ◽  
Takao Satoh ◽  
Takao Namihira ◽  
Hisato Saitoh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Atmospheric Pressure ◽  
Culture Medium ◽  
Plasma Jet ◽  
Helium Plasma ◽  
Cell Culture Medium
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