Journal: Stem Cell Research & Therapy

Article Title: A serum-free culture medium for long-term expansion of human airway basal stem cells

doi: 10.1186/s13287-026-04910-z

Figure Lengend Snippet: Evaluation of the effects of specific components in sfBSC medium on the growth of HBECs. A Brightfield images showing the morphology of HBECs at P3 and P4 after the removal of B27 supplement, GlutaMAX, L-ascorbic acid, Y27632, or EGF from the sfBSC medium. Complete sfBSC medium served as a control. Scale bar = 200 μm. B Quantitative assessment of HBEC proliferation using the CCK-8 assay following the removal of individual components (B27 supplement, GlutaMAX, L-ascorbic acid, Y27632, or EGF) from the sfBSC medium ( n = 3 per group). C Growth curves showing changes in cell numbers over time following the removal of rHSA, hydrocortisone or FGF10 from the sfBSC medium ( n = 3 per group). D Quantification of colony formation from single-cell seeding follwoing the removal of rHSA, hydrocortisone, or FGF10 ( n = 3 per group). E Brightfield images showing the morphology of HBECs at P3 to P6 after the removal of A-83-01, DMH1, or DAPT from the sfBSC medium. Complete sfBSC medium served as a control. Scale bar = 200 μm. White boxes highlight regions containing differentiated cells. F cPD curves for HBECs at different passages following removal of A-83-01, DMH1, or DAPT in sfBSC medium. Statistical significance at last day was determined by one-way ANOVA followed by Dunnett’s multiple comparisons test vs. complete medium; ***p < 0.001

Article Snippet: The complete sfBSC medium was supplemented with the following components: 1X B27 supplement (Gibco, 17504044), 0.1% recombinant human serum albumin (rHSA, Sigma-Aldrich, A9731), 2 mM Glutamax (Gibco, 35050061), 50 μg/mL L-ascorbic acid (Sigma-Aldrich, A4544), 0.4 μM monothioglycerol (MTG, Sigma-Aldrich, M6145), 1 μg/mL hydrocortisone (STEMCELL Technologies, 07926), 1 ng/mL recombinant human EGF (Sigma-Aldrich, SRP3027), 0.4 ng/mL recombinant human FGF10 (R&D systems, 345-FG), 10 μM Y27632 (Rho-associated protein kinase (ROCK) inhibitor, Selleck, S1049), 1 μM DAPT (γ-secretase/Notch signaling pathway inhibitor, Sigma-Aldrich, D5942), 1 μM A83-01 (ALK5 inhibitor, Sigma-Aldrich, SML0788), and 1 μM DMH-1 (BMP signaling inhibitor, Sigma-Aldrich, D8946).

Techniques: Control, CCK-8 Assay, Single Cell

Journal: Stem Cell Research & Therapy

Article Title: A serum-free culture medium for long-term expansion of human airway basal stem cells

doi: 10.1186/s13287-026-04910-z

Figure Lengend Snippet: Optimization of EGF and FGF10 concentrations for maintaining BSC morphology. A Brightfield images showing the morphology of HBECs cultured in media supplemented with different concentrations of EGF. Scale bar = 200 μm. White boxes demarcate cells exhibiting morphological alterations. B Brightfield images showing the morphology of HBECs cultured in media supplemented with different concentrations of FGF10. Scale bar = 200 μm. White boxes demarcate cells exhibiting morphological alterations. C Immunofluorescence staining of P63 and KRT5 in HBECs grown in medium supplemented with 40 ng/mL FGF10. Scale bar = 50 μm. D Immunofluorescence staining of NGFR in HBECs grown in media supplemented with 40 ng/mL FGF10. White borders demarcate regions with altered NGFR expression patterns. Scale bar = 50 μm

Article Snippet: The complete sfBSC medium was supplemented with the following components: 1X B27 supplement (Gibco, 17504044), 0.1% recombinant human serum albumin (rHSA, Sigma-Aldrich, A9731), 2 mM Glutamax (Gibco, 35050061), 50 μg/mL L-ascorbic acid (Sigma-Aldrich, A4544), 0.4 μM monothioglycerol (MTG, Sigma-Aldrich, M6145), 1 μg/mL hydrocortisone (STEMCELL Technologies, 07926), 1 ng/mL recombinant human EGF (Sigma-Aldrich, SRP3027), 0.4 ng/mL recombinant human FGF10 (R&D systems, 345-FG), 10 μM Y27632 (Rho-associated protein kinase (ROCK) inhibitor, Selleck, S1049), 1 μM DAPT (γ-secretase/Notch signaling pathway inhibitor, Sigma-Aldrich, D5942), 1 μM A83-01 (ALK5 inhibitor, Sigma-Aldrich, SML0788), and 1 μM DMH-1 (BMP signaling inhibitor, Sigma-Aldrich, D8946).

Techniques: Cell Culture, Immunofluorescence, Staining, Expressing