Fig. 1: Placental erythro-myeloid progenitors (PEMP) are exclusively found early in gestation. | Nature Communications

Fig. 1: Placental erythro-myeloid progenitors (PEMP) are exclusively found early in gestation.

From: Primitive haematopoiesis in the human placenta gives rise to macrophages with epigenetically silenced HLA-DR

Fig. 1

A Representative flow cytometry gating strategy identifying CD43+CD34+ placental erythro-myeloid progenitors (PEMP) in a 5 post-conception week (PCW) sample. B Flow cytometric expression of CD45, CD117 (c-Kit) and CD90 in PEMP, data are representative of n = 2 donors for CD90 expression and n = 3 donors for CD45 and CD117 expression. FMO fluorescence minus one. C Relative flow cytometry expression of HLA-A2 in PEMP, Hofbauer cells (HBC) and placenta-associated maternal macrophages (PAMM) from a sample with an HLA-A2 allotype mismatch between mother and foetus. D Representative Giemsa-stained cytospins of PEMP isolated by FACS. Scale bars, 20 μm. Representative images of 3 independent experiments from n = 3 donors. E Identification of CD34+ (grey) CD43+ (red) CD45+ (green) PEMP within the villous stroma of a 5 PCW placental sample. Scale bars, main panel 100 μm, inset panels 20 μm. Representative images of 3 independent experiments from n = 1 donor. F Flow cytometric analysis of PEMP abundance in a 5 PCW and a 9 PCW sample. G Quantification of PEMP frequency via flow cytometry as a proportion of all live cells, plotted against post-conception age in days, or (H) grouped by week. P value and R2 calculated via Pearson’s Correlation. Boxplot centre lines represent the median, with box limits showing the upper and lower quartiles, and whiskers denoting minimum and maximum values (n = 28 donors). I Principle component analysis embedding of flow cytometry data of HBC, PEMP and any phenotypic intermediates from 12 placental samples (gating strategy for analysis detailed in Fig. S1F–I). Cells were subjected to Phenograph clustering and clusters are coloured according to their cellular identities. n = 12 donors, 2 × 4 PCW, 2 × 5 PCW, 2 × 6 PCW, 2 × 7 PCW, 2 × 8 PCW and 2 × 9 PCW. PC principal component. J Mean expression of key flow cytometric markers (top panel), and distributions of phenograph clusters (bottom panel) along PC1. K PCA embeddings as in (I), with cells from each timepoint highlighted in blue within each panel.

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