Journal: bioRxiv
Article Title: Human ectodermal organoids reveal the cellular origin of DiGeorge Syndrome
doi: 10.1101/2025.08.08.669417
Figure Lengend Snippet: A) A heatmap showing differentially expressed genes that also include known markers for each respective differentiated NC derivative cell type at different timepoints of early and fully differentiated stages. B ) A 30-day protocol produces a pure population of pigmented MITF-immunopositive melanocytes that also express TYR and PMEL. MITF qPCR NC SD = ±0.002; Day 30 Melanocytes SD = ±4.919. MITF immuno percentage positive NC SD = 0; Day 30 Melanocytes SD = 0. TYR qPCR NC SD = ±0.008; Day 30 Melanocytes SD = ±223.7. PMEL immuno NC SD = ±0.021; Day 30 Melanocytes SD = ±0.6033. C ) Virtually all cells in the glial cultures differentiated into S100B-immunopositive cells. S100B qPCR NC SD = ±0.9851; D26 SD = ±2.476; D35 SD = ±2.853. S100B immuno NC SD = ±0.0001; D26 SD = ±0.0355; D35 SD = ±0.2323 D ) Virtually all sensory neurons express the cell type specific marker BRN3A. BRN3A qPCR NC SD = ±0.4091; D10 SD = ±20.05. BRN3A immuno D10 = ± 8.315, D16 = ±9.708. E ) Symphatetic neurons differentiated into a pure population of TH-immunopositive cells that also significantly increased expression of TBH at the end of the 16-day protocol. DBH qPCR NC SD = ±0.6501, D10 SD = ±0.7824, D16 SD = ±5.811. DBH imunno D10 SD = ±21.46, D16 SD = ±0. TH qPCR NC SD = ±0.3162, D10 SD = ±2.576, D16 SD = ±5.205. F ) Differentiation into chrommaffin cell lineage formed PNMT-positive cells in ten days. PNMT qPCR NC SD = ±0.1704, D10 SD = ±3.229. PNMT immuno Isotype SD = ±0.1115, D5 SD = ±0.1396, D10 SD = ±1.080. G ) Chondrocyte cultures formed Aggrecan and SOX9 -positive mineralizing chondrospheroids in 28 days. The cartilage lineage initiation and NC marker SOX9 expression is highest in early and midpoint cultures. SOX9 qPCR NC SD = ±0.4912, D14 SD = ±0.2429, D28 SD = ±0.07434. SOX9 immuno NC SD = ±0.1266, D14 SD = ±0.0939, D28 SD = ±0.1296. ACAN qPCR NC SD = ±0.0015, D14 SD = ±0.2845, D28 = ± 0.0596. ACAN immuno NC = ±0.0591, D14 SD = ±0.1144, D28 = ±0.1303. H ) In 28 days, NC cells organized into RUNX2-immunopositive osteocytes that have nuclei synchronized in a symmetrical angle. RUNX2 qPCR NC SD = ±0.0642, D14 SD = ±0.2367, D28 SD = ±0.3629. I ) Adipocytes express increasing levels of FABP4 during their 33 days on maturation resulting in a homogeneous culture of cells that form lipid-spot positive lipid droplets. FABP4 qPCR NC SD = ±0.0010, D11 SD = ± 0.0288, D33 SD = ±0.2864. LipidSpot immuno NC SD = ± 0.0942, D11 = ±0.1615, D33 = ±0.4709 J ) Transgelin and Smooth Muscle Actin expressing cells show a typical smooth muscle morphology after 21 days in a uniform culture. ACTA qPCR NC SD = ±1.269, D11 SD = ±2.676, D21 SD = ±5.413. ACTA immune NC SD = ±0.036, D11 SD = ±0.2735, D21 SD = ±0.4533. TAGLN qPCR NC SD = ±2.739, D11 SD = ±28.70, D21 = ±46.39. TAGLN immuno NC SD = ±0.0798, D11 SD = ±0.3547, D21 SD = ±0.5395. ISO= isotype IgG control (the images stained by additional antibodies are shown in ). Significance is annotated as such *: p-value > 0.05, **: p-value >0.002, ***: p-value >0.0002, ****: p-value >0.0001. NC: Neural Crest, Mel: Melanocytes, Sens Neu: Sensory Neurons, Symp Neu: Sympathetic Neurons, Chrom: Adrenal Chromaffin Cells, Chond: Chondrocytes, Osteo: Osteocytes, Adi: Adipocytes, Smo Mus: Smooth Muscle, hESC: human embryonic stem cells.
Article Snippet: H1 and H9 human embryonic stem cell (hESC) lines (WA09 (H9) and WA01 (H1) from WiCell, were used for the characterization of the ectodermal organoids.
Techniques: Marker, Expressing, Control, Staining
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