The mouse colon carcinoma cell line MC38, the Chinese hamster ovary cell line CHO-K1-hPD-L1, the CHO-K1-mPD-L1 line, and the human T lymphocytic leukemia Jurkat cell line were maintained in RMPI 1640 (Corning, NY, USA). All the cells were cultured in medium supplemented with 10% FBS (Biological Industries, Cromwell, CT, USA) and 100 U/mL penicillin/streptomycin (Solarbio, Beijing, China) in an incubator with 5% CO2 at 37 °C.
We conducted experiments employing CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells previously constructed in the laboratory. The flow cytometry results indicated high expression of PD-L1 in both CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells. Subsequently, we employed flow cytometry to assess the expression of PD-L1 in MC38 cells. The results revealed a high level of PD-L1 expression in MC38 cells. Also, the heightened PD-L1 expression in MC38 cells has been substantiated in multiple studies [32 (link),33 (link),34 (link)]. Notably, our experimental paradigm involved the stimulation of Jurkat cells with PHA and PMA, a methodology documented in the literature, known to elicit PD-1 overexpression within Jurkat cells [35 (link)].
We conducted experiments employing CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells previously constructed in the laboratory. The flow cytometry results indicated high expression of PD-L1 in both CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells. Subsequently, we employed flow cytometry to assess the expression of PD-L1 in MC38 cells. The results revealed a high level of PD-L1 expression in MC38 cells. Also, the heightened PD-L1 expression in MC38 cells has been substantiated in multiple studies [32 (link),33 (link),34 (link)]. Notably, our experimental paradigm involved the stimulation of Jurkat cells with PHA and PMA, a methodology documented in the literature, known to elicit PD-1 overexpression within Jurkat cells [35 (link)].
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