I have 3 questions regarding mixed lymphocyte reaction in mice:
1) I need to keep the mice alive (with intact spleen), so I want to use peripheral blood sample as a source of responder cells instead of splenocytes, is there any disadvantage in using this approach in mice (other than the lower yield)?
2) Is it necessary to perform T cell enrichment or negative selection for MLR responders or you can proceed with adjusting the responder:stimulator ratio assuming that T lymphocytes are 17-20 % ? (the target is proliferation assay only).
3) I came across a protocol for responder cell cryopreservation for primed MLR assay, however it is not detailed. Does anyone have experience with cryopreservation of lymphocytes for MLR assay?
Thank you,
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