RESUMO
Although the cell atlas of the human ocular anterior segment of the human eye was revealed by single-nucleus RNA sequencing, whether subtypes of lens stem/progenitor cells exist among epithelial cells and the molecular characteristics of cell differentiation of the human lens remain unclear. Single-cell RNA sequencing is a powerful tool to analyse the heterogeneity of tissues at the single cell level, leading to a better understanding of the processes of cell differentiation. By profiling 18,596 cells in human lens superficial tissue through single-cell sequencing, we identified two subtypes of lens epithelial cells that specifically expressed C8orf4 and ADAMTSL4 with distinct spatial localization, a new type of fibre cells located directly adjacent to the epithelium, and a subpopulation of ADAMTSL4+ cells that might be lens epithelial stem/progenitor cells. We also found two trajectories of lens epithelial cell differentiation and changes of some important genes during differentiation.
Assuntos
Cristalino , Humanos , Cristalino/metabolismo , Epitélio , Células Epiteliais/metabolismo , Olho , Diferenciação Celular , Análise de Sequência de RNARESUMO
OBJECTIVE: To clarify whether functionally competent dendritic cells (DC) can be generated from malignant pleural effusion in patients with lung cancer. METHODS: Malignant effusion-associated monocytes were separated by adherence from malignant effusion-associated mononuclear cells and cultured in medium with granulocyte macrophage colony-stimulating factor (GM-CSF) plus interleukin 4 (IL-4). TNF-alpha was added for the last 24 h before culture termination. Cultured DC were identified by (1) using microscopy, scanning electron microscopy, and immunocytochemistry for the morphological features; (2) phenotypic markers; and (3) functional characteristics including a high stimulatory capacity to activate proliferation of lymphocyte in an allogeneic mixed leukocyte reaction and the ability to produce high levels of IFN-gamma. RESULTS: Cultured DC had the typical morphological features. The phenotype of DCs generated from effusion showed higher expression of CD(86) (84.6 +/- 6.1)%, HLA-DR (81.1 +/- 13.0)%, CD(40) (42.0 +/- 21.7)%, CD(1a) (20.0 +/- 9.5)% and lower expression of CD(14) (4.8 +/- 3.5)% than the control group. There was a significant difference in the stimulatory activity in allogeneic lymphocyte proliferation and the ability to produce high levels of IFN-gamma between DC derived from the malignant effusion and the control group. CONCLUSION: These findings suggest that DC can be generated from malignant pleural effusion, which might be a useful source of DC for immunotherapy.