To establish the importance of fluorescein diacetate (FDA) as a viability stain for cultured hepatocytes. we hypothesized that FDA staining would correlate positively with hepatocyte viability and function. Mixtures of live and dead cells were stained with FDA and scanned by flow cytometry. A close correlation was observed between the live cell fraction and percent viability as determined by FDA staining (R2 = 0.962). Hepatocytes were also sorted into low fluorescence and high fluorescence groups. Both albumin production and lidocaine metabolism (P-450 activity) were significantly increased in the high fluorescence group compared to the low fluorescence group. An automated, fluorescence-activated assay was useful for rapid assessment of hepatocyte viability. In addition. the intensity of green fluorescence following staining with FDA correlated well with two specific measures of hepatocyte function.
Bibliographical noteFunding Information:
We thank Lenise E. Best and Michael A. Hupke for technical assistance with the flow cytometer and Prof. Friedrich Srienc for his helpful review of the data. This research was supported in part by grants from the Whittaker Foundation and the National Science Foundation (BCS- 89 15307).S LN was supported by an Ethicon/Society of University Surgeons Re- search Fellowship.
Copyright 2014 Elsevier B.V., All rights reserved.
- Albumin production
- Flow cytometry
- Fluorescein diacetate
- Lidocaine metabolism. viability