Shun Yu

This study describes a protocol for constructing human minor salivary gland (hMSG) organoids to establish a reproducible model for tissue regeneration research. Salivary gland stem/progenitor cells (hMSG-SCs) and mesenchymal stem cells (hMSG-MSCs) were isolated from pediatric minor salivary gland tissues, expanded, and co-seeded in a Matrigel-based three-dimensional (3D) system. Self-organization led to the formation of organoids with ductal-acinar-like structures, recapitulating native cellular heterogeneity more effectively than epithelial-only models. The current protocol emphasizes optimized culture conditions to preserve cell phenotypes and standardized steps for passaging, ratio mixing, and handling Matrigel (a commercial basement matrix), thereby enhancing reproducibility. Importantly, the presence of mesenchymal cells provides a supportive microenvironment that promotes epithelial survival, proliferation, and morphogenesis, overcoming limitations of low efficiency and incomplete structure. This approach enables consistent generation of salivary gland organoids and offers a robust model for studying development, functional restoration, and disease mechanisms, while also supporting potential translational applications in regenerative therapies.

This study describes a protocol for constructing human minor salivary gland (hMSG) organoids to establish a reproducible model for tissue regeneration research. Salivary gland stem/progenitor cells (hMSG-SCs) and mesenchymal stem cells (hMSG-MSCs) were isolated from pediatric minor salivary gland tissues, expanded, and co-seeded in a Matrigel-based three-dimensional (3D) system. Self-organization led to the formation of organoids with ductal-acinar-like structures, recapitulating native cellular heterogeneity more effectively than epithelial-only models. The current protocol emphasizes optimized culture conditions to preserve cell phenotypes and standardized steps for passaging, ratio mixing, and handling Matrigel (a commercial basement matrix), thereby enhancing reproducibility. Importantly, the presence of mesenchymal cells provides a supportive microenvironment that promotes epithelial survival, proliferation, and morphogenesis, overcoming limitations of low efficiency and incomplete structure. This approach enables consistent generation of salivary gland organoids and offers a robust model for studying development, functional restoration, and disease mechanisms, while also supporting potential translational applications in regenerative therapies.

To assess the association between sleep patterns and sleep factors (sleep duration, trouble sleeping, sleep disorder) and the risk of depression in older adults. A total of 5636 participants (2754 men and 2882 women) aged 60 years and older from the 2007–2014 waves of the National Health and Nutrition Examination Survey (NHANES) were included. Sleep duration, sleep problems, and sleep disorders were assessed in the home by trained interviewers using the Computer-Assisted Personal Interviewing (CAPI) system. The combined sleep behaviours were referred to as ‘sleep patterns (healthy, intermediate and poor)’, with a ‘healthy sleep pattern’ defined as sleeping 7–9 h per night with no self-reported trouble sleeping or sleep disorders. Intermediate and poor sleep patterns indicated 1 and 2–3 sleep problems, respectively. Baseline characteristics of participants analysed using one-way logistic regression. Logistic multiple linear regression was used to assess the association of sleep factors and sleep patterns with the risk of depressive symptoms.Conduct subgroup analyses to ensure robustness of findings. The overall prevalence of depression was 7.7% among the 5636 participants analysed, with the rate of depression in older women being 1.6 times higher than in older men. The prevalence of depression was higher in older adults with intermediate sleep pattern than in older adults with healthy sleep pattern (OR: 2.28, 95% CI: 1.71–3.03, p < 0.001). The prevalence of depression was higher in older adults with poor sleep pattern than in older adults with healthy sleep pattern (OR: 5.60, 95% CI: 4.25–7.39, p < 0.001). The findings were robust after controlling for sleep items in the PHQ-9. This nationally representative survey showed a relationship between sleep patterns and depression in older adults. However, the study population was limited to Americans, and we recommend continued investigation of the causal relationship and mechanisms between the two in the future, and further expansion of data sources in order to assess the applicability of the findings.