Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: The 3D culture of esophageal organoids in EEM hydrogel. a) Brightfield images of esophageal organoids cultured in various concentrations of EEM hydrogels and MAT at day 9. b) Quantification of organoid formation efficiency in EEM hydrogels, relative to MAT ( n = 3, one-way ANOVA with Tukey's multiple comparisons test; ∗∗ p < 0.01; ns, non-significant difference with p > 0.05). c) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogels and MAT at day 9 ( n = 4, one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). d) Relative gene expression of basal layer markers ( Sox2 and Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at day 9 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT). e) Morphology of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 9 days. f) Immunofluorescent staining for basal layer markers (CK14, SOX2, and p63), suprabasal layer markers (CK13 and CK4), F-actin, a cell-cell adhesion marker (ECAD), and a proliferation marker (Ki67) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days. g) Quantification of CK14 and CK13 positive area in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days, based on immunofluorescence-stained images ( n = 4; unpaired two-tailed t -test; ∗ p < 0.05 versus MAT; ns, non-significant difference with p > 0.05). h) Live/Dead staining to assess the viability of organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 11 days. i) Representative brightfield images at passage 0, 2, 4, and 6 of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT. Total culture days are given in parentheses. j) Comparison of gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at passage 0 and 6 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT at each passage).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Quantitative RT-PCR, Gene Expression, Marker, Two Tailed Test, Staining, Immunofluorescence, Comparison

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Neutral Buoyancy as a Simple Approach to Simulated Microgravity

doi: 10.1007/s13770-025-00781-2

Figure Lengend Snippet: The feasibility of the simulated microgravity in Optiprep with a neutral buoyancy regarding the stemness maintenance and trilineage differentiation of h BMSCs was evaluated. A qRT-PCR analysis and B western blotting determined the expression of OCT4, SOX2, and NANOG (stemness markers) in h BMSCs cultured at 3D-sim-μg (in Optiprep ), 3D-1 g (in CCM), and 2D-1 g (in CCM). C qRT-PCR analysis, D immunofluorescence staining, E histological analysis, and F quantitative analysis of osteogenic differentiation markers (RUNX2, OCN, and alizarin red S), adipogenic differentiation markers (PPARγ, CEBP/α, and Oil red O), and chondrogenic differentiation markers (SOX9, ACN, and Alcian blue) of h BMSCs cultured at 3D-sim-μg and 3D-1 g (n = 3; *p < 0.05 and **p < 0.01)

Article Snippet: The specific TaqMan Primers (Thermo Fisher Scientific) used were: OCT4 (Hs04260367_gH), SOX2 (Hs04234836_s1), NANOG (Hs02387400_g1), RUNX2 (Hs00231692_ml), osteocalcin (OCN; Hs00609452_gl), PPARγ (Hs01115513_m1), CEBP/α (Hs00269972_s1), SOX9 (Hs00165814_m1), aggrecan (ACN; Hs00153936_m1), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Hs02786624_g1), with GAPDH serving as the housekeeping gene.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Cell Culture, Immunofluorescence, Staining