title: Effective in vitro expansion of CD40-activated human B lymphocytes in a defined bovine protein-free medium.
abstract: CD40-CD154 interaction is used to culture human B lymphocytes, which are now viewed as effectors to potentially promote T lymphocyte response against malignant cells in cell-based therapy. Currently, the media used, based on bovine serum, are raising concerns for patient safety in such therapy. In this study, we have investigated whether human B lymphocytes could be cultured in the absence of bovine serum. Blood CD19(+) B lymphocytes were activated using interaction through CD40 in medium containing defined animals or human proteins and lipids, and were monitored during short-term periods (≤15 days). Conventional stem-cell medium and a medium containing human albumin instead of bovine albumin were tested. We observed that the response of B lymphocytes appeared influenced by lot-to-lot variability in low density lipoproteins (LDL). Nevertheless, B lymphocyte proliferation and secretion in serum-free and bovine protein-free media were quite similar to that of cells cultured in medium containing FBS. Our results show that CD40-activated B lymphocytes can be cultured for up to 15 days in a serum-free medium containing human albumin, LDL, α-tocopherol and chemically-defined lipids.
authors: Héma-Québec, Ingénierie cellulaire, Recherche et développement, Québec (Québec), Canada. sonia.neron@hema-quebec.qc.ca
authors: Sonia
authors: S
authors: Néron
authors: Annie
authors: A
authors: Roy
authors: Nellie
authors: N
authors: Dumont
authors: Nathalie
authors: N
authors: Dussault
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D019013:CD40 Antigens / Q000378:metabolism*; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D003470:Culture Media / Q000737:chemistry*; D016895:Culture Media, Serum-Free / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D008055:Lipids; D008077:Lipoproteins, LDL; D008213:Lymphocyte Activation; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D024502:alpha-Tocopherol
chemical_list: D019013:CD40 Antigens; D003470:Culture Media; D016895:Culture Media, Serum-Free; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008055:Lipids; D008077:Lipoproteins, LDL; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D024502:alpha-Tocopherol