Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium

Yabe, Noritsugu; Matsuya, Yutaka; Yamane, Isao; Takada, M

doi:10.1007/bf02623523

Cited by 29 publications

(14 citation statements)

References 18 publications

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“…Although the fusion method is well established for producing murine MoAbs, direct fusion of human MoAb-producing lymphocytes and cultured myeloma cells is rather difficult because hyperimmunization for P. aeruginosa does not occur during the course of natural infection. Further, human origin myeloma cell lines had lower fusion efficiency than that of murine origin (26).…”

Section: Discussionmentioning

confidence: 93%

Establishment of Stable Cell Lines Producing Anti‐Pseudomonas aeruginosa Monoclonal Antibodies and Their Protective Effects for the Infection in Mice

O'oka

Chonan

Mizutani

et al. 1992

Microbiology and Immunology

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Section: Discussionmentioning

confidence: 93%

Establishment of Stable Cell Lines Producing Anti‐Pseudomonas aeruginosa Monoclonal Antibodies and Their Protective Effects for the Infection in Mice

O'oka

Chonan

Mizutani

et al. 1992

Microbiology and Immunology

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“…We used two serum-free media to observe whether cells take up Dil-labeled lipid microspheres. ASF-301 medium (13) and SFM-101 (14) are mainly designed for culturing adherent cells and suspended cells, respective ly. The culture system is free from the effects of various substances such as lipoproteins, apolipoproteins and some proteins having opsonic effects.…”

Section: Discussionmentioning

confidence: 99%

Fluorescent Probe-Labeled Lipid Microsphere Uptake by Human Endothelial Cells: A Flow Cytometric Study

Suzuki¹,

Takahashi

Matsuki³

et al. 1992

Japanese Journal of Pharmacology

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Lipid microspheres have been used as carriers of drugs such as prostaglandin E, (lipo PGE1) and corticosteroid (liposteroid). Lipo-PGE, is used for the treatment of chronic arterial occlu sive diseases because its activity is far greater than that of free PGE, in vivo. To verify the fact that the drug carriers, lipid microspheres, are preferentially taken up by endothelial cells, we labeled lipid microspheres with a fluorescent probe, DiI (DiI-LM), and observed them in some in vitro models. Stoichiometric fluorescence was obtainable, and the fluorescence was stable between pH 3.3 and pH 8.9. Human umbilical vein endothelial cells and cells of a human endothelial cell line, ECV304, showed increased uptake of DiI-LM, 81% and 61%, respectively. In contrast, uptakes were less than 7% in hu man skin fibroblasts, 3T3 cells, and human neutrophils. Prominent perinuclear fluorescence was also observed in endothelial cells by fluorescence microscopy. DiI-LM and flow cytometric analysis will be useful for studies to elucidate the precise mechanism of the selective accumulation of lipid microspheres by cells in blood vessel walls.

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“…Murakami and co-workers reported ethanolamine as being essential for hybridoma growth under serum-free medium, and the ethanolamine in the GIT medium must also play an important role in hybridoma production (Murakami et al 1982;Kovar and Franek 1986). Yabe et al (1986) have described the development of a new serum-free medium, designated as NYSF which is a very peculiar and interesting culture medium for selective proliferation of B cell hybridomas. This NYSF medium is a 1:1 mixture of RPMI 1640 and MEM supplemented with hypoxanthine, thymidine, insulin, dihydroxyethylglycine, transferrin, ethanolamine, selenite, amino acids, and so on.…”

Section: Discussionmentioning

confidence: 99%

A great improvement of fusion efficiency in mouse B cell hybridoma production by use of the new culture medium, GIT.

Kudo

Morishita

Suzuki³

et al. 1987

Tohoku J. Exp. Med.

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The recently established culture medium, GIT, is applicable to many kinds of cells including mouse and human myeloma cells, and most adhesive cell lines. We applied this GIT medium to mouse B cell hybridoma production. When the medium was used to propagate myeloma cells before cell fusion and also for HAT selective medium, the fusion efficiency was more than twice as high as when the regular medium (RPMI-1640 supplemented with FBS) was used. Constantly more than 80% wells were hybridoma positive irrespective of the antigens used. To determine the optimal cell concentration at the hybridoma selection, a graded number of myeloma and spleen cells was distributed to each well ; the best result was obtained when 3 X 104 myeloma and 3 X 105 spleen cells were distributed to each well. In addition, the GIT medium shows very little lot-to-lot variation. These results indicate that fusion efficiency in mouse B cell hybridoma production was greatly improved by using GIT medium. hybridoma production ; GIT medium ; monoclonal antibody Since Kohler and Milstein (1975) introduced a new technology to obtain a monoclonal antibody directed against a single epitope on an antigen, monoclonal antibodies have been used extensively. The production of monoclonal antibodies to various antigens by the fusion of antigen-sensitized spleen cells with myeloma cells has been expanded to every biological field. Though basic methods have been well established, there is still a need for the more effective, stable fusion efficiency. Good and constant fusion efficiency makes it possible to effectively establish desired hybridoma clones and contributes markedly to the progress of

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Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium

Cited by 29 publications

References 18 publications

Establishment of Stable Cell Lines Producing Anti‐Pseudomonas aeruginosa Monoclonal Antibodies and Their Protective Effects for the Infection in Mice

Establishment of Stable Cell Lines Producing Anti‐Pseudomonas aeruginosa Monoclonal Antibodies and Their Protective Effects for the Infection in Mice

Fluorescent Probe-Labeled Lipid Microsphere Uptake by Human Endothelial Cells: A Flow Cytometric Study

A great improvement of fusion efficiency in mouse B cell hybridoma production by use of the new culture medium, GIT.

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