Decidual leukocytes play key roles in maternal-fetal tolerance and immunity. Here, we present detailed methods to purify, culture, and functionally analyze human placental dNK, dTreg, dTem, and dMɸ from decidua parietalis, the maternal part of the placental membranes; decidua basalis, the maternal part of the placenta; and placental villi. These sites have high clinical relevance in the development of villitis and chorioamnionitis. This allows in-depth phenotypic and functional investigation of placental immune populations and their interactions with extravillous trophoblasts.

For complete details on the use and execution of this protocol, please refer to Ikumi et al.,¹ Tilburgs et al.,² Salvany-Celades et al.,³ Crespo et al.,⁴ van der Zwan et al.⁵