图1 iPSC衍生的心脏类器官构建策略[31]

Fig. 1 Strategy for constructing cardiac organoids derived from iPSC[31] (a) Formation scheme of cardiac organoids; (b) Cardiac organoids forming three layers; (c) Fluorescent staining of myocardial marker MHC; (d) Staining images of cTnT and WT1; (e) Staining image of mesenchymal cell marker vimentin CHIR: laduviglusib (CHIR99021); MHC: myosin heavy chain; cTnT: cardiac troponin T; WT1: Wilms’ tumor 1

图2 LCS生物陶瓷调控NSCs的行为[51]

Fig. 2 LCS bioceramics regulating the behaviors of NSCs[51] (a) Schematic diagram of preparation and application of neural platform based on LCS; (b) SEM images of LCS; (c) Representative immunofluorescence staining images of Nestin and MAP2 expression in NSCs; (d) Representative immunofluorescence staining images of GFAP and Tuj1 protein expression in NSCs; (e) Representative dual immunofluorescence staining images of GFAP and Tuj1 in NSCs under different culture conditions; (f) Representative immunofluorescence staining images of matural neuron marker MAP. MAP2: microtubule-associated protein 2; GFAP: glial fibrillary acidic protein; Tuj1: neuronal class III β-tubulin clone

图3 MS纳米球调控内皮细胞和真皮细胞相互作用[55]

Fig. 3 MS nanospheres regulating the interaction between endothelial cells and dermal cells[55] (a) TEM image of MS nanospheres; (b) EDS mappings of elements including Mg, Si and O in MS nanospheres; (c) Fluorescence image displaying the biomimetic spatial distributions of human umbilical vascular endothelial cells (HUVECs, red fluorescence) and human hair dermal papilla cells (HHDPCs, green fluorescence); (d) Stained images of cell clusters during co-culture; (e, f) Expression of genes related to angiogenesis (e) and hair follicle formation (f) in co-culture and single culture systems; (g) Microscopic observation of hair growth in newborn skin area of nude mice; (h) Distribution of mature hair follicles in co-culture group. VEGF: vascular endothelial growth factor; HIF-1α: hypoxia-inducible factor 1-α; KDR: kinase insert domain receptor; VE-cad: Vascular endothelial cadherin; PDGF-α: platelet-derived growth factor receptor α; PDGF-β: platelet-derived growth factor receptor β; c-Myc: cellular myelocytomatosis oncogene; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

图4 CS纳米线释放的生物活性离子用于促进肠道类器官发育[47]

Fig. 4 Bioactive ions released by CS nanowires for promoting intestinal organoid development[47] (a) Proliferation of intestinal organoids treated with CS nanowires of different concentrations in culture medium; (b, c) Gene expression of four differentiated cell types treated with 0, 50 and 100 μg/mL CS nanowires for 3 (b) and 5 days (c); (d) Immunofluorescence staining of β-catenin in intestinal organoids treated with CS nanowires for 3 and 5 days; (e) Heat maps of gene expression related to Wnt/β-catenin signaling pathway in intestinal organoids after 3 and 5 days of CS nanowire treatment; (f) Relative glucose absorption of different groups; (g) Relative ATP production of different groups. Wnt3: Wingless-type MMTV integration site family, member 3; LRP6: Low-density lipoprotein receptor-related protein 6; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

图5 CNT促进肠道类器官发育[61]

Fig. 5 CNT for promoting intestinal organoid development[61] (a) Typical images of epithelial subpopulations in intestinal organoids treated with CNT (50 μg/mL) at different time periods; (b) Representative SEM images showing degradation of the matrix over time during development of intestinal organoids; (c) Statistical analysis of changes over time in phosphorylated p38 mitogen activated protein kinase (P-p38 MAPK), p38 mitogen activated protein kinase (p38 MAPK), yes-related protein (YAP), and phosphorylated yes-related protein (P-YAP) in intestinal organoids treated with CNT (50 μg/mL); (d) Quantification of YAP nuclear translocation over time in intestinal organoids treated with CNT (50 μg/mL); (e) Representative confocal images of mitochondrial membrane potential and relative mitochondrial activity in intestinal organoids over time; (f) Effects of different amino acids on the proliferation and differentiation of intestinal organoids; (g) Proposed mechanism of CNT induced intestinal organoid development. SWCNTs: single-walled carbon nanotubes; MWCNTs: multiwalled carbon nanotubes; L-FABP: liver fatty acid-binding protein; ISC: intestinal stem cell; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

图6 肠道类器官在探究口服生物玻璃微球减少肠道炎症损伤中的应用[71]

Fig. 6 Application of organoids in exploring reduction of intestinal inflammatory damage by bioactive glass microspheres[71] (a) Concentration of TNF-α in macrophage culture medium after different treatments; (b) Expression of anti-inflammatory gene ARG in macrophage culture after different treatments; (c) CCK-8 testing of intestinal organoids cultured under different media conditions; (d) Immunofluorescence staining of Ki67 in intestinal organoids cultured under different conditions for 144 h; (e) Morphology and size of intestinal organoids cultured under different conditions for 72 and 144 h. BG: bioactive glass; LPS: lipopolysaccharide; CK: non-treated control; ARG: arginase; Conditioned media were collected from untreated RAW “CM-RAW-CK”, LPS-activated RAW “CM-LPS”, and LPS-activated RAW after pretreatment of BG extract liquids (1 : 100 dilution ratio) “CM-LPS/BG”; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively