Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells

Figure 3

Comparison of the purity of hepatic stellate cells isolated via iohexol gradient without or with subsequent cell sorting. Purity of hepatic stellate cells (HSC) before (left top) and after fluorescence-activated cell sorting (FACS, right top) (a) based on their retinol autofluorescence only (dashed line) or additional exclusion of HSC-Kupffer cell doublets (highly pure real HSC, straight line). The statistical summary of retinol⁺ cells compared to HSC (with HSC-KC doublets excluded) is depicted (b). Analysis of purity using cell type-specific markers for major cell populations. Decorin is considered as a marker for hepatic stellate cells, C-type lectin domain family 4f (Clec4f) is a gene expressed by Kupffer cells, the platelet/endothelial cell adhesion molecule 1 (Pecam-1, CD31) is mainly expressed by endothelial cells, and albumin is a marker for hepatocytes (c). Data are given relative to the expression of β-actin of the cells derived from perfusion and digestion. Mean data ± SD of independent experiments.

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