Expansion of isolated B cells with the B Cell Expansion Kit, human

This application protocol describes the in vitro expansion of primary human B cells.  
Many downstream applications require expansion of B cells, due to their low frequencies in peripheral blood. In vitro cultivation of B cells can be time consuming and can often result in dead cells and low expansion rates. The B Cell Expansion Kit, human has a defined formulation that enables standardized expansion of isolated B cells. Expanded cells are fully functional and ready for downstream applications. The expansion rate reaches up to a 10-fold increase after 14 days of in vitro culture. The phenotype and characteristics of the expanded B cells are also comparable to the starting material, therefore preserving all B cell subsets. 


Isolation of human PBMCs

Reagent preparation 

Preparation of PEB buffer (for staining of isolated B cells)

PEB buffer: Prepare a solution containing phosphate-buffered saline (PBS), pH 7.2, 0.5% bovine serum albumin (BSA), and 2 mM EDTA by diluting MACS® BSA Stock Solution (# 130-091-376) 1:20 with autoMACS® Rinsing Solution (# 130-091-222). Keep buffer cold (2–8 °C). Always use freshly prepared buffer. 
Do not use autoMACS Running Buffer or MACSQuant® Running Buffer as they contain a small amount of sodium azide that could affect the results.


The following is a listing of reagents, instruments, and consumables needed for each step of this protocol. These products are for research use only.
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