I am doing mixed lymphocyte reaction for testing the reaction of C57BL/6 splenocytes to Balb/c splenocytes. I cultured the whole splenocytes of Balb/c mice with the irradiated splenocytes of C57/BL6 mice to be used as APCs. I tested for the proliferation of cells using both CFSE and MTT assay. Although I can analyze some proliferation of cells in the form of small  peaks using CFSE method but my MTT assay did not work. I did not observe any formazon crystal formation after 5-7 days of culture. Can anybody suggest me what can be the reason for failure for my MTT assay although it worked for my positive control samples (samples stimulated with anti-CD3)?

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