Journal: Stem Cell Reviews and Reports

Article Title: Reenacting Neuroectodermal Exposure of Hematopoietic Progenitors Enables Scalable Production of Cryopreservable iPSC-Derived Human Microglia

doi: 10.1007/s12015-022-10433-w

Figure Lengend Snippet: RNA sequencing analysis reveals close transcriptional similarity between primary human microglia and iPSdMiG. A PCA showing that iPSdMiG from all three lines (with n = 3 technical replicates per line) cluster in closest proximity to primary human microglia (n = 38 independent samples; in silico data) and comparatively farther away from iMGL (n = 9 independent samples; in silico data), THP1 macrophages (n = 6 technical replicates), iPSCs (n = 3 technical replicates) and cortex samples (n = 16 independent samples; in silico data). B TF enrichment analysis based on the gene core signature of iPSdMiG and primary human microglia (|log 2 FC|≥ 3 and FDR-adjusted p-value ≤ 0.001) identified a highly similar set of TFs driving cellular identity in both cell populations. Green circled genes were common in both primary human microglia and iPSdMiG. Pale orange circled genes were upregulated in iPSdMiG, while pale blue circled genes were upregulated in primary human microglia. C Heatmap of lineage-specific genes (microglial, pluripotency and neuronal genes) demonstrating that little to no pluripotency and neuronal gene transcripts are detectable in iPSdMiG and primary human microglia, whereas microglial genes are highly expressed in both cell types. D WGCNA identified a subset of genes (module five; dotted-black line) that were co-expressed in iPSdMiG, primary human microglia and iMGL. E Pathway enrichment analysis showed that module five primarily consisted of genes associated with innate immune pathways

Article Snippet: Once firmly adhered, EB-loaded macrocarriers were transferred into suspension culture formats such as T75 non-tissue culture-coated flasks and placed on a rocker within a humidified incubator for further differentiation in expansion media composed of STEMdiff TM APEL TM 2 (Stem Cell Technologies, Vancouver, Canada), 10% knock-out serum replacement (KSR; Gibco), 1 × N2 supplement, 1 × B27 supplement, 25 ng/ml rh interleukin (IL) 3 (rhIL3), 25 ng/ml rhIL34 and 25 ng/ml rh macrophage colony stimulating factor (rhMCSF; all from Peprotech).

Techniques: RNA Sequencing, In Silico