Abstract
Albumin, transferrin, and lipids can replace serum entirely for support of LPS-stimulated murine B lymphocytes in culture. In the presence of these compounds, growth and maturation to IgM and IgG secretion, induced by lipopolysaccharide (LPS), occurs at the same or higher efficiency in serum-free conditions as in conventional serum-containing medium, even at relatively low cell concentrations. In contrast to the rapid disappearance of LPS reactivity in conventional serum-containing medium, responsiveness remains at initial levels in serum-free conditions for 2 days before slowly declining. Overall lymphocyte survival is also markedly prolonged. In the presence of thymus "filler" cells, the serum-free conditions permit growth of every LPS-responsive cell to a clone of Ig-secreting cells at dilutions as low as a single reactive B cell per culture. The results have several important implications. These include the establishment for the first time of transferrin as a requirement for B lymphocyte responses in culture, and the availability now of conditions for the assay isolation of cell products regulating lymphocyte function, free of interference from undefined serum components.
MeSH Terms
Albumins
Animals
B-Lymphocytes/immunology,physiology
Blood
Cells, Cultured
Culture Media
Immunoglobulin G/metabolism
Immunoglobulin M/metabolism
Lipids
Lipopolysaccharides/pharmacology
Mice
Rats
Transferrin
Chemicals
Albumins
Culture Media
Immunoglobulin G
Immunoglobulin M
Lipids
Lipopolysaccharides
Transferrin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Iscove N N
Melchers F
References (25)
25 references, click to expand
-
Role of bovine serum albumin in blastoid transformation of lymphocytes by phytohemagglutinin.
Proc Soc Exp Biol Med. 1977 Mar;154(3):444-8
PMID: 847004
-
Leakage of sucrose from phosphatidylcholine liposomes induced by interaction with serum albumin.
Biochim Biophys Acta. 1977 Mar 29;497(1):183-91
PMID: 849477
-
Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte.
Cell. 1977 Jan;10(1):27-34
PMID: 319912
-
Frequencies of mitogen-reactive B cells in the mouse. I. Distribution in different lymphoid organs from different inbred strains of mice at different ages.
J Exp Med. 1977 Jun 1;145(6):1511-9
PMID: 301174
-
Development of methods for the quantitative in vitro analysis of androgen-dependent and autonomous Shionogi carcinoma 115 cells.
Cell. 1977 Jan;10(1):35-44
PMID: 837442
-
A plaque assay for all cells secreting Ig of a given type or class.
Eur J Immunol. 1976 Aug;6(8):588-90
PMID: 789096
-
Interaction of phosphatidylcholine with bovine serum albumin. Specificity and properties of the complexes.
Biochim Biophys Acta. 1976 Mar 18;427(1):325-36
PMID: 944054
-
IgM in bone marrow-derived lymphocytes. Synthesis, surface deposition, turnover and carbohydrate composition in unstimulated mouse B cells.
Eur J Immunol. 1974 Mar;4(3):170-80
PMID: 4604559
-
The mitogenic effect of lipopolysaccharide on bone marrow-derived mouse lymphocytes. Lipid A as the mitogenic part of the molecule.
J Exp Med. 1973 Apr 1;137(4):943-53
PMID: 4571328
-
Identification of albumin as the serum factor essential for the growth of activated human lymphocytes.
J Biol Chem. 1976 Feb 25;251(4):987-92
PMID: 2610
-
The kinectics of proliferation and maturation of mitogen-activated bone marrow-derived lymphocytes.
Eur J Immunol. 1974 Oct;4(10):687-91
PMID: 4547710
-
Induction of immunoglobulin and antibody synthesis in vitro by lipopolysaccharides.
Eur J Immunol. 1972 Aug;2(4):349-53
PMID: 4563347
-
Modification of red cell membrane structure by cholesterol-rich lipid dispersions. A model for the primary spur cell defect.
J Clin Invest. 1975 Jan;55(1):115-26
PMID: 162782
-
Structure and function of transferrins. I. Physical, chemical, and iron-binding properties.
Arzneimittelforschung. 1974 Apr;24(4):476-85
PMID: 4408363
-
Promotion of replication in lymphoid cells by specific thiols and disulfides in vitro. Effects on mouse lymphoma cells in comparison with splenic lymphocytes.
J Exp Med. 1973 Sep 1;138(3):574-92
PMID: 4727914
-
Identification of transferrin as a lymphocyte growth promoter in human serum.
Exp Cell Res. 1972 Sep;74(1):163-9
PMID: 4672470
-
Enhancement of antibody synthesis in vitro by mercaptoethanol.
Cell Immunol. 1972 Jan;3(1):156-60
PMID: 5061825
-
Replacement of serum by hormones permits growth of cells in a defined medium.
Nature. 1976 Jan 15;259(5539):132-4
PMID: 813153
-
Modification of Escherichia coli membranes in the prereplicative phase of phage T4 infection. Specificity of association and quantitation of bound phage proteins.
J Biol Chem. 1975 Mar 25;250(6):2339-50
PMID: 803972
-
Role of mercaptoethanol and endotoxin in stimulating B lymphocyte colony formation in vitro.
J Immunol. 1976 Mar;116(3):635-8
PMID: 1082889
-
Growth of B-lymphocytes clones in semisolid culture is mitogen dependent.
J Exp Med. 1976 May 1;143(5):1265-70
PMID: 1083421
-
Continuous growth of mitogen-reactive B lymphocytes.
Scand J Immunol. 1975;4(8):853-8
PMID: 1082636
-
Selenium is an essential trace nutrient for growth of WI-38 diploid human fibroblasts.
Proc Natl Acad Sci U S A. 1976 Jun;73(6):2023-7
PMID: 1064872
-
Stimulation of DNA synthesis in cultures of mouse spleen cell suspensions by bovine transferrin.
Exp Cell Res. 1969 Feb;54(2):195-200
PMID: 5776472
-
Removal of fatty acids from serum albumin by charcoal treatment.
J Biol Chem. 1967 Jan 25;242(2):173-81
PMID: 6016603