Kupffer Cells Promote the Differentiation of Adult Liver Hematopoietic Stem and Progenitor Cells into Lymphocytes via ICAM-1 and LFA-1 Interaction
Kupffer cells promote liver hematopoiesis via the interaction between ICAM-1 and LFA-1. (a) The expression of ICAM-1 and VCAM-1 on kupffer cells from PBS- (blue) or LPS-treated mice (orange) was analyzed by flow cytometry. (b) Flow cytometry was used to measure the expression of LFA-1 and VLA-4 on liver LSK cells from PBS- or LPS-treated mice. (c) Immunofluorescence staining shows CFSE-labeled LSK cells (green) in close contact with ICAM-1+ (blue) kupffer cells (F4/80+, red) in the liver. Bar: 20 μm. (d) Freshly isolated kupffer cells were preincubated with anti-ICAM-1 (10 μg/mL) and subsequently cocultured with sorted liver CFSE-labeled LSK cells in the presence of anti-ICAM-1 for 24 h. Nonadherent CFSE+ cells were counted by flow cytometry and compared with those from cocultures in the absence of anti-ICAM-1. (e) Lin- cells isolated from the liver were cocultured with freshly isolated kupffer cells in the presence of anti-ICAM-1 in vitro. The proportion of LSKs in the coculture system was detected by flow cytometry on day 7. (f) Statistical analysis of the absolute number of LSK cells and Lin- cells. (g) Freshly isolated LSK cells were cocultured with kupffer cells in the presence of PBS or anti-ICAM-1. The cocultured cells were collected every seven days for analysis by flow cytometry. The statistical percentage of lymphocytes CD3+ T and CD19+ B cells in the LSK-kupffer cell coculture system. Three replicate wells were established for each group. Data are represented as the .;.
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