Stempro cell dissociation reagent

Leukocytes were isolated from human decidual tissue (Table S2) as previously described (Xu et al., 2015 ). Briefly, the decidua basalis was collected from the basal plate of the placenta, and the decidua parietalis was separated from the chorioamniotic membranes. Decidual tissue was homogenized using a gentleMACS Dissociator (Miltenyi Biotec, San Diego, CA, USA) in StemPro Cell Dissociation Reagent (Life Technologies, Grand Island, NY, USA). Homogenized tissues were incubated for 45 min at 37°C with gentle agitation. After incubation, tissues were washed with 1X PBS (Life Technologies) and filtered through a 100 μm cell strainer. Cell suspensions were collected and centrifuged at 300 × g for 10 min at 4°C, and the cell pellet was suspended in stain buffer (BD Biosciences, San Jose, CA, USA). Mononuclear cells were purified using a density gradient (Ficoll-Paque Plus; GE Healthcare Bio-Sciences, Up-psala, Sweden), following the manufacturer’s instructions. Then, mononuclear cell suspensions were washed using stain buffer prior to immunophenotyping.

Decidual leukocytes from human decidual tissue were isolated as previously described (73 (link)). Briefly, the decidua basalis was collected from the basal plate of the placenta, and the decidua parietalis was separated from the chorioamniotic membranes. Decidual tissue was homogenized using a gentleMACS Dissociator (Miltenyi Biotec, San Diego, CA, USA) in StemPro Cell Dissociation Reagent (Life Technologies, Grand Island, NY, USA). Homogenized tissues were incubated for 45 min at 37°C with gentle agitation. After incubation, tissues were washed with ice-cold 1X PBS and filtered through a 100 µm cell strainer. Cell suspensions were collected and centrifuged at 300 x g for 10 min at 4ºC, and the cell pellet was suspended in stain buffer (Cat#554656; BD Biosciences, San Jose, CA, USA). Mononuclear cells were purified using a density gradient (Ficoll-Paque Plus; GE Healthcare Bio-Sciences, Uppsala, Sweden), following the manufacturer’s instructions. Lastly, mononuclear cell suspensions were washed using stain buffer before immunophenotyping.