Journal: Cell Stem Cell

Article Title: Generation of human appetite-regulating neurons and tanycytes from pluripotent stem cells

doi: 10.1016/j.stem.2026.05.005

Figure Lengend Snippet: Signaling factors required for generation of human ARC in vitro (A) Hypothalamic development in vivo and in vitro differentiation of hPSC into hypothalamic cells. (B) Principal-component analysis (PCA) of day 16 RT-qPCR data (RC17), clustering according to ARC- and PVN-associated genes. Each dot represents one differentiation batch ( n = 113). (C) Estimated marginal means assessing the effect of factors on PC1, adjusted for covariates. (D) ICC/ISH at days 16 and 50 for ARC protocol with or without BMP4. Scale bars: 100 μm. (E) ARC differentiation protocol and maturation options. (F–H) ICC of day 50 in 2D (F) and 3D (G and H), scale bars: 50 and 100 μm. (I) Quantification of day 50 3D spheroids (AGRP: n = 14, 4.53% ± 0.69%, mean ± SEM; αMSH: n = 6, 5.90% ± 1.19%). (J) Xenotransplantation of day 16 ARC progenitors into nude rats. (J′) Top: ICC for human neural cell adhesion molecule (NCAM) highlights graft site, scale bar: 2 mm. Bottom: IF for human nuclear antigen (HuNu), scale bar: 200 μm. (K–Q) IF for ARC neuronal markers in grafts. Scale bars: 100 μm (K and O), 50 μm (N), 25 μm (L, M, P, and Q).

Article Snippet: BMP4 , Miltenyi Biotec , Cat#130-111-168.

Techniques: In Vitro, In Vivo, Quantitative RT-PCR

Journal: Cell Stem Cell

Article Title: Generation of human appetite-regulating neurons and tanycytes from pluripotent stem cells

doi: 10.1016/j.stem.2026.05.005

Figure Lengend Snippet: Duration of BMP stimulation controls ARC versus VMH identity (A) BMP signaling in the developing chick tuberal hypothalamus. Adapted from Chinnaiya et al. vFB, ventral forebrain. (B) Experimental design to simulate BMP withdrawal. (C) ICC/ISH for days 16 and 50 across BMP4 withdrawal conditions. Scale bar: 100 μm. (D) Day 16 RT-qPCR (RC17, KOLF2.1J, BIONi010) across BMP4 withdrawal conditions. Mean ± SEM, Kruskal-Wallis with Dunn’s multiple comparisons test: FGF10 : −BMP versus days 5–14 p < 0.0001; days 5–7 versus days 5–14 p < 0.0001; days 5–9 versus days 5–14 p = 0.0040; days 5–11 versus days 5–14 p = 0.013; SOX14 : −BMP versus days 5–14 p < 0.0001; days 5–7 versus days 5–14 p = 0.0004; days 5–9 versus days 5–14 p = 0.0011; days 5–11 versus days 5–14 p = 0.0020. (E) Day 50 RT-qPCR (RC17, KOLF2.1J, BIONi010) across BMP4 withdrawal conditions. Mean ± SEM, Kruskal-Wallis with Dunn’s test: AGRP : −BMP versus days 5–14 p = 0.0080; days 5–7 versus days 5–14 p = 0.0269 NR5A1 : −BMP versus days 5–14 p = 0.0016; days 5–7 versus days 5–14 p = 0.0162. (F and G) UMAPs of day 16 (F) and days 50 + 80 + 110 (G) snRNA-seq ( n = 3; RC17, KOLF2.1J, BIONi010) and annotated clusters with proportional representation. ARC- and VMH-specific clusters are marked with a box. (H) Spatial projection probability of days 50 + 80 + 110 clusters onto human adult reference. (I) 3D staining for Wisteria floribunda agglutinin (WFA), scale bar: 100 μm.

Article Snippet: BMP4 , Miltenyi Biotec , Cat#130-111-168.

Techniques: Quantitative RT-PCR, Staining

Journal: Cell Stem Cell

Article Title: Generation of human appetite-regulating neurons and tanycytes from pluripotent stem cells

doi: 10.1016/j.stem.2026.05.005

Figure Lengend Snippet: Different POMC subtypes are dependent on BMP timing (A) UMAP of days 50 + 80 + 110 snRNA-seq data from G with POMC clusters and their % across BMP4 exposure timings. (B) Key marker genes across POMC + ARC and VMH clusters in in vitro dataset and human fetal and adult reference datasets. , , (C) MetaNeighbor cluster similarity analysis comparing in vitro POMC⁺ clusters to hypothalamic reference datasets. , , (D) Expression of ARC/VMH-related markers on spatial transcriptomics reference. (E) DEG between in vitro POMC⁺ subtypes, spatial DEGs from HYPOMAP are highlighted. Benjamini-Hochberg corrected p values. (F) Prediction probabilities from spatial mapping of in vitro POMC clusters to HYPOMAP. (G) Hybridization chain reaction (HCR) of chick embryo heads at HH17 and HH20, n = 7–10. Scale bar: 100 μm.

Article Snippet: BMP4 , Miltenyi Biotec , Cat#130-111-168.

Techniques: Marker, In Vitro, Expressing, Spatial Transcriptomics, Hybridization