I am interested in learning of preferred formulations of tissue culture media used to grow human B cells (CD19 cells) isolated from peripheral blood mononuclear cells (PBMCs) of Leukopaks using commercial B cell extraction kits.
If possible, I want to keep B cells growing and healthy for at least six days. During the incubation period, I would need to extract partial amounts of B cells for analysis and transferring to new media for continued growth.
A formulation I recently used is presented below. It was sufficient to keep cells in high enough quantity and viability for a period of four days after extraction.
B Cell Base Medium Version 6
RPMI 1640 1X with L-Glutamine
Fetal Bovine Serum (FBS), Heat-Inactivated In-House, 10% Final Concentration
Penicillin-Streptomycin Solution, 2% Final Concentration
Sodium Pyruvate Solution
Gibco GlutaMAX Solution
MEM Non-Essential Amino Acids (NEAA) Solution
HEPES Liquid Solution
Human Recombinant Insulin Zinc Solution
2-Mercaptoethanol
B Cell Base Medium Version 6 Supplements (Added at Time of Use)
ODN 2006 Class B CpG Oligonucleotide Human TL9 Ligand
Recombinant Human Interleukin 2
Recombinant Human Interleukin 4
Recombinant Human Interleukin 21
Purified Anti-Human CD40 Antibody
Do supplements such as Interleukins need to be at higher concentrations for B cells compared to T cells and other immunological cells?
Is use of commercially-prepared tissue culture media kits specific for B cells from companies such as StemCellTechnologies and Miltenyi Biotec a preferred approach instead of preparing an in-house tissue culture medium?
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