Protocol using lentivirus to establish THP-1 suspension cell lines for immunostaining and confocal microscopy
Wangsheng Ji1, Lianfei Zhang2, Xinqi Liu2
1State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Protein Science, Frontiers Science Center for Cell Responses, Department of Biochemistry and Molecular Biology, College of Life Sciences, Nankai University, Tianjin 300071, China; Joint National Laboratory for Antibody Drug Engineering, the First Affiliated Hospital of Henan University, Henan University, Kaifeng 475000, China.
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THP-1, a monocyte cell line growing in suspension, is widely used in immunology research. However, establishing suspension cell lines and performing confocal microscopy can be challenging. Here, we present a protocol to efficiently generate THP-1 cell lines using lentivirus and perform immunostaining and confocal microscopy. We detail steps for virus production, THP-1 cell infection and clone selection, fixing the suspension cells to the glass slide for immunostaining, and subsequent confocal microscopy. This protocol can be applied to other suspension cells. For complete details on the use and execution of this protocol, please refer to Ji et al. (2021).1.
