Establishment of long-term serum-free culture for lacrimal gland stem cells aiming at lacrimal gland repair

摘要

Isolation and characterization of lacrimal gland stem cells (LGSGs). The strategy of LGSG primary and continuous passage culture. , The morphology of LGSCs in primary culture at day 7; scale bar, 200 μm, 100 μm. Hematoxylin and eosin staining of LGSCs at day 7; scale bar, 50 μm. Immunofluorescent staining of LGSCs at day 7. LGSCs express epithelial cell marker E-cadherin (red), scale bar, 50 μm; LGSCs express stem cell marker Krt14 (red), scale bar, 50 μm; LGSCs express proliferative cell marker Ki67 (red), scale bar, 50 μm. Counterstain, DAPI (4′,6-diamidino-2-phenylindole, blue). The morphology of LGSCs in continuous passage culture. LGSGs cultured for 1, 3, 5, and 7 days, scale bar, 100 μm; LGSCs cultured in different passages (P1, P10, P20, and P40), scale bar, 100 μm. The cell numbers of LGSCs cultured for 7 days in different passages (P1, P10, P20, and P40). The cells were seeded at 1 × 10 cells per well (24-well plate). The experiments were performed three times and the average cell number is shown in the graph. Transcriptional expression of adult stem/progenitor cell markers of the mouse LG and different passage LGSCs (P1, P10, P20, and P40). LG, lacrimal gland; NC, negative control