The isolated T cells will be further activated and expanded in vitro for transduction (with chimeric antigen receptors). The ex vivo culturing will take about 2-3 weeks.
I know that for isolating rare cell populations (I.e. pDCs) it is recommended to pool spleens together to obtain sufficient numbers but not sure if the same can be done for mouse T cells. Would there be any concerns of cross-reactivity despite the cells coming from the same background (C57Bl6 in our case) only different individuals?
When isolating mouse T cells, it is generally recommended to use spleens from individual mice rather than pooling them together. This is because T cell populations can vary among individuals, even within the same strain, due to genetic and environmental factors. Pooling spleens from different mice may introduce variability in the T cell populations, including differences in T cell receptor repertoires and activation states.
To obtain sufficient numbers of T cells for downstream applications, you can isolate T cells from multiple individual spleens separately and then combine the isolated T cells together after purification. This approach ensures that the T cell populations remain representative of each individual mouse, minimizing the potential for cross-reactivity or other unwanted effects.
By isolating T cells from individual spleens, you can also control the quality and purity of the T cell population, ensuring consistency in your experiments. Additionally, if any issues arise during the ex vivo culturing process, it allows for easier troubleshooting and identification of potential sources of variability.
In summary, it is generally recommended to isolate T cells from individual mouse spleens rather than pooling them together to maintain consistency and minimize potential issues related to cross-reactivity or variability in T cell populations.
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