构建稳定表达PDL1的B16F10细胞株

Construction of a B16F10 cell line that stably express PDL1

目的通过转染及流式细胞仪(flow cytometry,FCM)筛选出稳定表达小鼠细胞程式死亡配体1(Programmed cell death ligand 1,PDL1)的B16F10细胞株。方法将携带PDL1的真核表达质粒通过脂质体法转染到B16F10细胞株中,经FCM进行多次分选,筛选出稳定表达PDL1的B16F10细胞株,将其与抗CD3/抗CD28抗体活化的CD4^+T细胞共培养48 h后,FCM检测CD4^+T细胞表面CD69以及CD25的表达。结果建立了稳定表达PDL1的B16F10细胞株,该细胞株表面的PDL1可诱导CD4^+T细胞高表达CD25。结论成功地得到了稳定表达PDL1的B16F10细胞株,为后续研究奠定了物质基础。

Objective To establish and screen a stable B16F10 cell line expressing mouse programmed cell death ligand 1(PDL1)via transfection and flow cytometry(FCM).Methods The plasmid containing PDL1 cDNA was transfected into B16F10 cell lines using jetPEITM DNA transfection reagent.B16F10 cell lines that stable express PDL1 were obtained by multiple sorting by FCM.The anti-CD3/anti-CD28 activated CD4+T cells were co-cultured with B16F10 cells for 48 hours and then the CD69 and CD25 expression on CD4+T cells were detected by flow cytometry.Results Stable transfected B16F10 cell line expressing PDL1 was established and the PDL1on the surface of B16F10 cell lines activated CD4+T cells to express high levels of CD25.Conclusions The B16F10 cell line that stable expression of PDL1 was successfully obtained and it provided solid experimental foundation for further studies.