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Non enzymatic liver cells isolation: citrate-perchlorate technique
M C Camberos1, C L Gomez Dumm, V D'Alessandro
1CEDIE, Hospital de Pediatría Dr. Ricardo Gutiérrez, La Plata, Argentina.
Acta Physiologica, Pharmacologica Et Therapeutica Latinoamericana : Organo De La Asociacion Latinoamericana De Ciencias Fisiologicas Y [De] La Asociacion Latinoamericana De Farmacologia
|January 1, 1996
Summary
This study introduces a new non-enzymatic method to isolate rat hepatocytes. The technique yields metabolically and morphologically normal hepatocytes, confirmed by electron microscopy.
Area of Science:
- Hepatology
- Cell Biology
- Biochemistry
Background:
- Hepatocyte isolation is crucial for research.
- Existing enzymatic methods can alter cell function.
- A non-enzymatic approach is needed for preserving hepatocyte integrity.
Purpose of the Study:
- To develop and validate a non-enzymatic method for isolating rat hepatocytes.
- To assess the metabolic and morphological integrity of isolated hepatocytes.
Main Methods:
- Rat liver perfusion via the portal vein using a specialized non-enzymatic solution.
- Purification of hepatocytes using a diatrizoate density gradient.
- Morphological assessment via electron microscopy.
Main Results:
- Successful isolation of rat hepatocytes without enzymatic treatment.
- Electron microscopy confirmed well-preserved hepatocytes with normal ultrastructure.
- Isolated hepatocytes demonstrated metabolic and morphological normality.
Conclusions:
- The described non-enzymatic perfusion technique effectively isolates viable rat hepatocytes.
- This method provides a reliable alternative for obtaining metabolically and morphologically intact hepatocytes for research.