Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells

Figure 4

Cultures of hepatic stellate cells isolated via iohexol density gradient centrifugation without or with subsequent cell sorting. Hepatic stellate cells (HSC) were isolated from 40-week-old C57BL/6J mice using enzymatic digestion of the liver based on pronase and collagenase, followed by density gradient centrifugation in 8% iohexol. Cells were cultured directly after the gradient for one day (a) and four days (b), where Kupffer cells (indicated by arrows) can be found in the HSC culture. Highly pure HSC after additional fluorescence-activated cell sorting (FACS) after one (c) and four days of culture are shown (d). Expression analysis of desmin (e) and phalloidin (f) of HSC after four days of culture, indicating proper maturation of HSC.

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