Journal: Bioengineering & Translational Medicine

Article Title: Fostering cell–cell interactions and integrating angiocrine factors to promote the development of salivary microtissues in 3D

doi: 10.1002/btm2.70118

Figure Lengend Snippet: Media screening for 2D maintenance of hS/PCs and HUVECs. (a) Proliferation of hS/PCs (i) and HUVECs (ii) under different media conditions (HEP: 100% HepatoSTIM, HEP + EGM2: 50/50 (v/v) mixture of HepatoSTIM and EGM2, EGM2: 100% EGM2). Data is normalized to the initial number of cells seeded on day 0. Error bars represent SEM, n = 3. * p < 0.05, ** p < 0.01, one way ANOVA post Hoc Tukey test. (b) Representative confocal images of hS/PCs immunofluorescently stained for keratin 5 (K5, i) and keratin 14 (K14, ii) after 5 days of culture in different media. K5/K14: Green, nuclei: Blue, F‐Actin: Red. Scale bar: 50 μm. (C) Representative confocal images of HUVECs immunofluorescently stained for VE‐cadherin (i) and CD31 (ii) after 5 days of culture in different media. VE‐cadherin/CD31: Green, nuclei: Blue, F‐Actin: Red. Scale bar: 50 μm.

Article Snippet: Here, soluble factors in EGM2 acted as angiocrine signals, contributing to the growth and maturation of the epithelial spheroids.

Techniques: Staining

Journal: Bioengineering & Translational Medicine

Article Title: Fostering cell–cell interactions and integrating angiocrine factors to promote the development of salivary microtissues in 3D

doi: 10.1002/btm2.70118

Figure Lengend Snippet: Characterization of HUVEC monoculture maintained on HA hydrogels in EGM2. (a) Representative confocal images of HUVECs stained with calcein AM (green‐live cells) and ethidium homodimer‐1 (red‐dead cells) after 2, 7, and 14 days of culture. White circles indicate dead cells. Scale bar = 100 μm. (b) Percent viable HUVECs as a function of culture time. ns = non‐significant. (c) qPCR analyses of HUVEC‐specific markers across different time points, normalized to day 2. n = 3. Error bars represent SEM, * p < 0.05, ** p < 0.01, *** p < 0.005, Housekeeping gene: GAPDH , one way ANOVA with Post Hoc Tukey test as the reference gene. (d–g) Characterization of HUVEC monocultures by immunofluorescence. CD31, VE‐cadherin, Vimentin and vWF: Green, nuclei: Blue, F‐Actin: Red. Scale bar = 20 μm.

Article Snippet: Here, soluble factors in EGM2 acted as angiocrine signals, contributing to the growth and maturation of the epithelial spheroids.

Techniques: Staining, Immunofluorescence

Journal: Bioengineering & Translational Medicine

Article Title: Fostering cell–cell interactions and integrating angiocrine factors to promote the development of salivary microtissues in 3D

doi: 10.1002/btm2.70118

Figure Lengend Snippet: Characterization of hS/PC monocultures maintained in HA gels in EGM2. (a) Representative confocal images of hS/PC spheroids stained with calcein AM (green‐live cells) and ethidium homodimer‐1 (red‐dead cells) after 1, 7, and 14 days of culture. Scale bar = 100 μm. (b) Percent viable cells in hydrogel‐derived hS/PC spheroids as a function of culture time. ns, non‐significant. (c) 3D rendering of a representative confocal image of a complex hS/PC microtissues established in HA gels on day 14. Nuclei: Magenta and F‐Actin: Green. x , y , z dimensions are 300, 300, 30 μm, respectively. (d–h) qPCR analyses of cellular expression of stem/progenitor markers (d, e), differentiation markers (f, g), and ECM, cell–cell, and cell–ECM adhesion proteins (h) across different time points normalized to the day 2 of culture. n = 3. Error bars represent SEM, * p < 0.05, ** p < 0.01, ** p < 0.001, housekeeping gene: GAPDH , one way ANOVA with Post Hoc Tukey test.

Article Snippet: Here, soluble factors in EGM2 acted as angiocrine signals, contributing to the growth and maturation of the epithelial spheroids.

Techniques: Staining, Derivative Assay, Expressing

Journal: Bioengineering & Translational Medicine

Article Title: Fostering cell–cell interactions and integrating angiocrine factors to promote the development of salivary microtissues in 3D

doi: 10.1002/btm2.70118

Figure Lengend Snippet: Morphological and phenotypic characterization of cocultured HUVECs and hS/PCs. (a) Schematic representation of the coculture configuration. The culture was maintained in EMG2. (b) Representative confocal images of HUVEC monolayer and hS/PC microtissue established on day 14 on/in HA gels with EGM2. Nuclei: Blue, F‐Actin: Red. Scale bar = 50 μm. (c–f) qPCR analyses of hS/PC expression of epithelial cell‐relevant markers and proteins across different time points, normalized to day 2. (g, h) qPCR analyses of HUVEC expression of endothelial cell‐relevant markers and proteins across different time points, normalized to day 2. (i, j) qPCR analyses of hS/PC expression of epithelial cell‐relevant markers and proteins in epithelial‐endothelial cocultures, normalized to the corresponding epithelial monocultures on day 14. (k) qPCR analyses of hUVEC expression of endothelial cell‐relevant markers and proteins in epithelial‐endothelial cocultures, normalized to the corresponding endothelial monocultures on day 14. For all qPCR analyses, n = 3. Error bars represent SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, housekeeping gene: GAPDH , one‐way ANOVA with Post Hoc Tukey test or Student's t ‐test.

Article Snippet: Here, soluble factors in EGM2 acted as angiocrine signals, contributing to the growth and maturation of the epithelial spheroids.

Techniques: Expressing