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R J Westerwoudt

Publications and source records attributed to R J Westerwoudt.

5 recordsLinked to original sources
PubMed1984

Improved fusion technique. II. Stability and purity of hybrid clones.

Optimal conditions are defined for hybridoma formation between mouse spleen cells and mouse myeloma cells. The results of using different numbers of spleen cells in the fusion process is reported in 2 parts. Part I deals with the number of spleen cells in relation to hybridoma formation and antibody production. Part II treats of the purity of hybridoma clones and the loss of antibody production following fusion. Part I. Two series of experiments show that when cell fusion is performed properly the total number of antibody producing clones is greater than in non-standard conditions. The yield of hybridomas obtained with a ratio of mouse myeloma to mouse spleen cells of 1:10 did not differ from that reported by De Blas et al. (1981). The number of hybridomas formed seems to depend mainly on the number of mouse spleen cells available. The most satisfactory yield of monoclonal antibodies is obtained under conditions producing growth in approximately 100% of the wells. Part II. Two weeks after fusion a number of antibody producing clones were cultured in limiting dilution. Analysis of the hybridomas indicated that at least 40% of the antibody producing clones disappear during the first 3 weeks. Antibody producing hybridomas were as a rule not outgrown by non-antibody producing clones.

Animals↗
PubMed1983

Improved fusion technique. III. The growth promoting activity of lipopolysaccharide, dextran sulfate, and red cell lysate added to Hy-clone calf serum.

A new high quality young-calf serum, Hy-clone calf serum (HcCS), was tested for use in hybridoma culture. This calf serum alone had little growth promoting activity and was much inferior to fetal calf serum (FCS). Red cell lysate (RCL) used in combination with the young-calf serum showed very good growth promoting activity. Growth was increased about threefold over that in the presence of FCS. However, HcCS and RCL could not substitute for feeder cells when hybridomas were cultured as single cells under conditions of limiting dilution. It is thought likely that the potent growth promoting factor in red cell lysate is hemoglobin.

Animals↗
PubMed1983

Improved fusion technique. I. Human umbilical cord serum, a new and potent growth promoter, compared with other B cell and hybridoma activators.

Accelerated proliferation of hybridoma cells was observed in the presence of human umbilical cord serum (HUCS). This had very strong growth-promoting activity, even at a concentration of 2%. A comparison was made between HUCS and other B cell growth promoters, such as lipopolysaccharide (LPS) and dextran sulfate (DxS), macrophage supernatant, and human endothelial culture supernatant (HECS). The growth-promoting effect of HUCS was superior. Using a microcytotoxicity assay, we found no significant differences in the number of antibody producing clones with the various culture media, except for fetal calf serum.

Animals↗