The use of .BETA.-cyclodextrin in lymphocyte culture. Induction of the primary antibody response in vitro in calf serum- or newborn bovine serum-containing culture medium.

Abstract

In order to induce the primary antibody response to sheep erythrocytes in vitro in murine lymphocytes, the cells are usually cultured in RPMI-1640 medium containing 10% fetal calf serum (FCS). When FCS was replaced by calf serum (CS) or newborn bovine serum (NBS), the antibody response was poor compared with that obtained with FCS. However, a comparable response was induced when CS or NBS was added at 1% to the culture medium in combination with 500 micrograms/ml beta-cyclodextrin (beta-CD). In the presence of beta-CD, the response was the highest when CS or NBS was added at 1%. An important observation was that all the lots of CS (5 lots) and NBS (2 lots) tested supported the response equally in the presence of beta-CD. The anti-sheep erythrocytes response induced by 1% CS or NBS in the presence of beta-CD was shown to be completely dependent on the added antigen, suggesting that beta-CD is not a polyclonal activator. beta-CD showed neither mitogenic nor cytotoxic effects on murine lymphocytes. These results indicate that beta-CD is a useful additive for inducing the primary antibody response in vitro in the culture medium containing CS or NBS which is usually less supportive but less expensive than FCS.

Demonstration of the Requirements for Cholesterol or Low Density Lipoprotein in the Antibody Response of Murine Lymphocytes in Serum-free Culture

“…Elution was done with n-hexane-isopropyl alcohol (98: 2), and the elution profiles were recorded at 210nm. I, authentic cholesterol;11, spot C component; III, authentic cholesterol plus spot C component. of authentic cholesterol, suggesting that the major active component in FCS ex.…”