Establishment of Normal Cell-Derived Monocyte Cell Line Stably Expressing the ZsGreen1 Gene

Abstract: It is demanded in the field of immunology to establish monocyte cell lines derived from normal human cells that stably express exogenous genes. To address this question， we employed multiple methods to transfect the ZsGreen1 gene into the normal human SC monocytic cells， including Lipofectamine® 3000， adenovirus and Tet-on element containing lentivirus. We found that the transfection/infection rates by using Lipofectamine® 3000 and adenovirus in SC cells were 0.66% and 0%， respectively. The infection rate of Tet-on lentivirus was 8.5% in SC cells. ZsGreen1+ SC cells (SC-ZsGreen1 cells) were then screened out and cultured， resulting in the acquisition of two SC-ZsGreen1 cell strains. Both stains reached over 95% of the positive ZsGreen1 expression rate. Next， we used PMA to allow these cells to differentiate into macrophages. In addition to the upregulated phagocytotic effect， PMA introduced increased CD11b and CD14 expression， and promoted IL-8 secretion in both SC-ZsGreen1 cell stains， similar to SC cells. In conclusion， we reported a feasible strategy for the establishment of normal cell-derived monocyte cell lines stably expressing the exogenous gene of ZsGreen1.

CSTR: 32200.14.cjcb.2018.10.0002