(A) Schematic representation of the derivation of trophoblast organoids (TOs) and decidual organoids (DOs) from mid-to-late gestation placental tissue. (B) Representative bright-field images of TOs …
(A) Immunohistochemistry in human full-term placental villi tissue used to generate TOs. Shown are no primary controls (left) and representative images of tissue immunostained for cytokeratin-19 …
Shown is a three-dimensional image reconstruction of an xz-stack of TOs immunostained for SDC-1 (in green) and counterstained for actin (in red). DAPI-stained nuclei are shown in blue. A single xz …
(A) Volcano plot demonstrating differentially expressed transcripts in TOs or DOs as assessed by DeSeq2 analysis. Gray circles represent genes whose expression was not significantly changed and red …
(A, B) Principal component analysis based on transcriptional profiles from DOs (orange) or TOs (blue). (C) Transcripts on the first and second principal components (PC1, PC2) with the greatest …
(A, B) Heatmap of 12 conditioned medium (CM) preparations generated from four established TO (A) and DO (B) lines analyzed by Luminex-based multianalyte profiling for the indicated cytokines, …
Concentrations of IL-6 (blue) and IFN-λ2 (red) in CM generated from a single TO line throughout the first 25 days of culture.
(A, B) Comparison of the levels of the type I IFNs IFN-α2 and IFN-β and type III IFNs IFN-λ1 and IFN-λ2 in conditioned medium (CM) isolated from TOs (A) or DOs (B) as determined by Luminex assays. (C…
(A, B) Volcano plots demonstrating differentially expressed transcripts in TOs (A) or DOs (B) treated with poly I:C as assessed by DeSeq2 analysis. Gray circles represent genes whose expression was …
(A) Venn diagram denoting unique and shared differentially expressed transcripts in TOs (blue circle) or DOs (orange circle) treated with poly I:C. (B) Heatmap (based on log2 RPKM values) of …
(A, B) Representative confocal micrographs of DOs (A) or TOs (B) infected with human CMV (HCMV) strains EGFP-tagged AD169r and mCherry-tagged TB40E for ~48 hr. Mock infected shown in top row of (A) …
(A, B) Volcano plots demonstrating differentially expressed transcripts in TOs (A) or DOs (B) infected with HCMV strains AD169r or TB40E (as denoted at top) by DeSeq2 analysis. Gray circles …
(A, B) Heatmaps demonstrating the induction of factors at left (shown as fold change from mock-treated controls) from TOs (A) and DOs (B) infected with HCMV (AD169r, top or TB40E, bottom strains). …
(A) Black and white images of single channel confocal micrographs used to generate the merged images shown in Figure 8I. (B) Representative over the counter (OTC) pregnancy tests for hCG in two …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Other | Matrigel | Corning | 356231 | Growth-factor-reduced for organoid growth. |
Other | TrypLE Express | Gibco | 12605–028 | For organoid dissociation during passaging. |
Other | Stem Pro Accutase | Gibco | A11105-01 | For organoid dissociation during passaging. |
Other | CryoStor CS10 stem cell freezing medium | STEMCell Technologies | 07930 | For organoid cryopreservation. |
Other | Trypsin-250 | Alfa Aesar | Alfa Aesar | For stem cell isolation from tissue. |
Other | Collagenase V | Sigma-Aldrich | C9263-100MG | For stem cell isolation from tissue. |
Other | recombinant human EGF | Gibco | PHG0314 | For organoid growth media. |
Other | recombinant human R-spondin 1 | R & D systems | 4645-RS-100 | For organoid growth media. |
Other | recombinant human FGF2 | Peprotech | 100–18 C | For organoid growth media. |
Other | recombinant human HGF | Peprotech | 100–39 | For organoid growth media. |
Other | ITS-X supplement | Gibco | 51500–056 | For organoid growth media. |
Other | KO serum replacement | ThermoFisher | 10828010 | For organoid growth media. |
Other | NRG1 | Cell Signaling | 5218SC | For organoid growth media. |
Other | A83-01 | Tocris | 2939 | For organoid growth media. |
Other | Y27632 | Sigma or STEMCell Technologies | Y0503 or 72304 | For organoid growth media. |
Other | nicotinamide | Sigma | N0636 | For organoid growth media. |
Other | FBS | Cytiva HyClone | SH30070.03 | For organoid growth media. Heat inactivated at 50 oC for 30 min. |
Other | Advanced DMEM/F12 medium | Life Technologies | 12634–010 | For organoid growth media. |
Other | 24-well tissue culture plates | Corning | 3526 | For organoid plating. |
Cell line (human) | HEK-Blue IFN-λ reporter cells | InvivoGen | hkb-ifnl | |
Other | HCMV AD169r BADrUL131-Y4 | T. Shenk | PMID: 16051825 | |
Other | HCMV TB40/E | N. Moorman | PMID: 21307184 | |
Other | 8-well chamber slide | Millipore | PEZGS0816 | For organoid plating. |
Other | Cell recovery solution | Corning | 354253 | |
Other | Vectashield | Vector Laboratories | H-1200–10 | Mounting medium for immunofluorescence. |
Antibody | Mouse monoclonal Cytokeratin-19 | Abcam | ab9221 | (Immunofluorescence, 1:500) |
Antibody | Rabbit monoclonal Cytokeratin-19 | Abcam | ab52625 | (Immunofluorescence, 1:500) |
Antibody | Rabbit polyclonal E-cadherin | Invitrogen | PA5-85088 | (Immunofluorescence, 1:500) |
Antibody | Rabbit monoclonal EpCAM | Abcam | ab223582 | (Immunofluorescence, 1:500) |
Antibody | Mouse monoclonal Mucin 5AC | Abcam | ab3649 | (Immunofluorescence, 1:500) |
Antibody | Rabbit polyclonal SIGLEC6 | Abcam | ab262851 | (Immunofluorescence, 1:500) |
Antibody | Mouse monoclonal HLA-G | Abcam | ab52454 | (Immunofluorescence, 1:500) |
Antibody | Rabbit monoclonal HLA-G | Abcam | ab283260 | (Immunofluorescence, 1:500) |
Antibody | Mouse monoclonal Ki67 | BD Biosciences | 550609 | (Immunofluorescence, 1:500; Immunohistochemistry, 1:200) |
Antibody | Rabbit monoclonal SDC-1 | Abcam | ab128936 | (Immunofluorescence, 1:500) |
Other | Alexa Fluor 594–conjugated phalloidin | Invitrogen | A12381 | For actin staining. (Immunofluorescence 1:1000) |
Other | Alexa Fluor 633–conjugated phalloidin | Invitrogen | A22284 | For actin staining. (Immunofluorescence 1:1000) |
Other | Alexa Fluor Plus 488 Goat anti-Mouse IgG secondary antibody | Invitrogen | A32723 | Secondary antibody for immunofluorescence. (Immunofluorescence 1:1000) |
Other | Alexa Fluor 594 Goat anti-Mouse IgG secondary antibody | Invitrogen | A11032 | Secondary antibody for immunofluorescence. (Immunofluorescence 1:1000) |
Other | Alexa Fluor 488 Goat anti-Rabbit IgG secondary antibody | Invitrogen | A11034 | Secondary antibody for immunofluorescence. (Immunofluorescence 1:1000) |
Other | Alexa Fluor 594 Goat anti-Rabbit IgG secondary antibody | Invitrogen | A11037 | Secondary antibody for immunofluorescence. (Immunofluorescence 1:1000) |
Software, algorithm | Fiji | N/A | https://imagej.net/software/fiji/ | For image analysis, including three- dimensional reconstructions. |
Software, algorithm | CellSens | Olympus | N/A | |
Software, algorithm | Imaris | Oxford Instruments | Version 9.2.; for image analysis, including three- dimensional reconstructions. | |
Software, algorithm | CLC Genomics | Qiagen | Version 20 | |
Software, algorithm | GraphPad Prism | Dotmatics | Version 9 | |
Other | Vectastain Elite ABC HRP Kit | Vector Laboratories | PK-6100 | For immunohistochemistry. |
Other | Biotinylated Goat Anti-Rabbit | Vector Laboratories | BA-1000 | For immunohistochemistry. (Immunohistochemistry, 1:250) |
Other | Biotinylated Goat Anti-Mouse | Vector Laboratories | BA-9200 | For immunohistochemistry. (Immunohistochemistry, 1:250) |
Other | DAB substrate | Vector Laboratories | SK-4100 | For immunohistochemistry. |
Other | Vectamount Permanent Mounting Medium | Vector Laboratories | H-5000 | For immunohistochemistry. |
Commercial assay, kit | GenElute total mammalian RNA miniprep kit | Sigma | RTN70-1KT | With on-column DNAse treatment |
Commercial assay, kit | iScript cDNA synthesis kit | Biorad | 1708891 | N/A |
Commercial assay, kit | iQ SYBR Green Supermix | Biorad | 1708882 | N/A |
Software, algorithm | DeSeq2 | M.Love et al | PMID: 25516281 | https://bioconductor.org/packages/release/bioc/html/DESeq2.html |
Software, algorithm | pheatmap | Raivo Kolde | https://cran.r-project.org/web/packages/pheatmap/pheatmap.pdf | |
Software, algorithm | pcaExplorer | Marini and Binder | PMID: 31195976 | https://bioconductor.org/packages/release/bioc/html/pcaExplorer.html |
Commercial assay, kit | Bio-Plex Pro Human Inflammation Panel 1 | Biorad | 171AL001M | |
Commercial assay, kit | Bio-Plex Human Chemokine Panel, 40-plex | Biorad | 171AK99MR2 | |
Commercial assay, kit | Bio-Plex Pro Human Cytokine Screening Panel, 48-Plex kit | Biorad | 12007283 | |
Commercial assay, kit | hCG Human ProcartaPlex Simplex Kit | Thermo Fisher | EPX010-12388-901 | |
Commercial assay, kit | CA125/Mucin-16 Human ProcartaPlex Simplex Kit | Thermo Fisher | EPX010-12437-901 | |
Chemical compound, drug | Poly(I:C) HMW | Invivogen | tlrl-pic | |
Chemical compound, drug | Ruxolitinib | Invivogen | tlrl-rux | |
Biological sample | Primary human trophoblasts from full term placentas | PMID: 27066743 | SRP067137 | Bulk RNASeq from primary human trophoblasts. |
Biological sample | Primary human trophoblasts from full term placentas | PMID: 28784796 | SRP109039 | Bulk RNASeq from primary human trophoblasts. |
Biological sample | First trimester placental tissue | University of Cambridge | PRJEB38810 | Bulk RNASeq from first trimester placental tissue. |
Biological sample | Full-term placental tissue | PMID: 33969332 | PRJNA721668 | Bulk RNASeq from ffull-term placental tissue. |