What question did this study set out to answer?

To establish and evaluate an enzymatic perfusion and density gradient centrifugation protocol for isolating and subfractionating intact adult rat hepatocytes.

July 1, 1975The Journal of Cell BiologyOpen Access

Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions.

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Authors

PDP DrochmansUniversité Libre de Bruxelles JWJean‐Claude WansonUniversité Libre de Bruxelles RMR MosselmansUniversité Libre de Bruxelles

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Implication

Experimental study demonstrates separation of distinct adult rat hepatocyte subpopulations via Ficoll gradient centrifugation, suggesting zonal segregation into centrolobular and perilobular...

Key Points

To establish and evaluate an enzymatic perfusion and density gradient centrifugation protocol for isolating and subfractionating intact adult rat hepatocytes.
Perfused adult rat livers with calcium-free Hanks' solution followed by collagenase and hyaluronidase enzymatic digestion and gentle mechanical rolling to release single cells.
Separated the isolated parenchymal cell suspension using isopycnic density gradient centrifugation on Ficoll gradients.
Assessed cell recovery, cell diameters, ultrastructural preservation, glucose-6-phosphatase activity, and yields of protein, RNA, DNA, and glycogen.
Yielded 60% to 65% of liver parenchymal cells with 70% recovery of protein, DNA, and RNA relative to liver homogenates, with preserved ultrastructure and unaltered glucose-6-phosphatase activity.
Resolved two distinct hepatocyte subpopulations: light hepatocytes (mean diameter 20.5 µm, mean density 1.10) with extensive smooth endoplasmic reticulum, and heavy hepatocytes (mean diameter 19.0 µm, mean density 1.14) characterized by dense glycogen accumulations.
Demonstrated structural and morphological differences between the fractions, indicating enrichment of centrolobular cells in the light fraction and perilobular cells in the heavy fraction.

Cite This Study

Drochmans et al. (1975) studied this question.

synapsesocial.com/papers/6a6f5fc2c92390ac2d079a95 https://doi.org/10.1083/jcb.66.1.1

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Implication

Experimental study demonstrates separation of distinct adult rat hepatocyte subpopulations via Ficoll gradient centrifugation, suggesting zonal segregation into centrolobular and perilobular...

Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions.

Discussion

Member takes

Implication

Key Points

Cite This Study

Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions.

Discussion

Member takes

Implication

Key Points

Cite This Study

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