Isolation and Labeling of B Cells

Spleens were isolated and pressed through a 70-μm nylon screen to generate a single-cell suspension. B cells were isolated using the EasySep Mouse B cell kit (StemCell Technologies) per the manufacturer’s protocols. CTV or CFSE labeling was performed following the manufacturer’s protocols. 5 × 10⁶ purified B cells were injected into each lobe of the thymus using ultrasound guidance as previously described (59 ).

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Martinez R.J., Breed E.R., Worota Y., Ashby K.M., Vobořil M., Mathes T., Salgado O.C., O’Connor C.H., Kotenko S.V, & Hogquist K.A. (2023). Type III interferon drives thymic B cell activation and regulatory T cell generation. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2220120120.

Publication 2023

B cells Cell Cfse Mouse Nylon Stemcell Thymus Ultrasound

Corresponding Organization :

Other organizations : University of Minnesota Medical Center, Rutgers, The State University of New Jersey

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Variable analysis

independent variables

Isolation of spleens
Pressing spleens through a 70-μm nylon screen
B cell isolation using EasySep Mouse B cell kit
CTV or CFSE labeling of purified B cells

dependent variables

Generation of a single-cell suspension
Percentage/number of B cells after isolation

control variables

Manufacturer's protocols for B cell isolation and CTV/CFSE labeling
Ultrasound guidance for B cell injection into the thymus

controls

No positive or negative controls were explicitly mentioned in the provided information.

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