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Analytical methods for the study of liver cell proliferation.
P Gerlyng1, T Stokke, H S Huitfeldt
1Department of Tissue Culture, Norwegian Radium Hospital, Oslo.
Cytometry
|January 1, 1992
Summary
Microscopy and image cytometry are effective for analyzing rat liver regeneration, measuring DNA synthesis and cell division. Flow cytometry offers speed but cannot differentiate cell types, while microscopy provides detailed analysis.
Area of Science:
- Hepatology
- Cell Biology
- Biotechnology
Background:
- Regenerating liver growth analysis requires accurate measurement of DNA synthesis, ploidy, and binucleation.
- Existing cytometric methods present limitations in comprehensively analyzing these parameters.
Purpose of the Study:
- To evaluate various cytometric methods for analyzing regenerating rat liver growth.
- To compare the efficacy of microscopy, flow cytometry, and image cytometry for assessing hepatocellular DNA ploidy, binucleation, and DNA synthesis.
Main Methods:
- Microscopic counting of immunostained hepatocytes for bromodeoxyuridine (BrdUrd) incorporation.
- Flow cytometry analysis of BrdUrd and DNA-stained cells.
- Image cytometry using fluorescence or Feulgen staining.
Main Results:
- Microscopy allows measurement of overall growth rate, binucleation rate, and indices for mononuclear and binuclear cells.
- Flow cytometry provides labeling indices for DNA ploidy classes and nonparenchymal cells but cannot distinguish mononuclear from binuclear hepatocytes.
- Image cytometry, though slower, enables complete analysis of DNA ploidy and nuclearity, making it suitable for cell cultures.
Conclusions:
- Image cytometry is a potentially superior method for analyzing regenerating liver, especially in cell cultures.
- Fluorescence staining is preferred over Feulgen staining for simultaneous analysis with BrdUrd.
- A non-immunological method is useful for tumor analysis where cell dissociation is challenging.