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It is thirty years since the technique of high-yield preparation of
isolated hepatocytes, by collagenase perfusion of the liver, was
published. The original method described by Berry and Friend has
undergone many minor modifications by other workers, and the
two-step procedure introduced by Seglen in 1976 has become the most
frequent way to prepare hepatocyte suspensions. An important
development introduced by Bissell in 1973 was the use of the cells
as the first step in monolayer culture. The availability of the
isolated hepatocyte preparation as cells in suspension or culture
has undoubtedly facilitated research on the liver. This was
emphasised in our book, published (with Dr. Greg Barritt) in 1990,
which described in detail methods of preparation and the properties
of the isolated hepatocytes. It also discussed the usefulness of
the preparation for the study of intermediary and xenobiotic
metabolism, calcium ion transport, and the growth and
differentiation of hepatocytes in culture. The book also touched
briefly on a range of specialised techniques, including peri
fusion, subcellular fractionation, transplantation,
cryopreservation and measurement of intracellular pH. Although
standard procedures for the manipulation of hepatocytes have not
changed a great deal in ten years, they have undoubtedly been
refined. This applies particularly to hepatocyte culture
techniques, cryopreservation, and even to preparation of hepatocyte
suspensions, where it is now feasible to use purified enzymes.
There is also much more emphasis on the use and study of human
hepatocytes, particularly in the field of pharmacology and
therapeutics.
It is thirty years since the technique of high-yield preparation of
isolated hepatocytes, by collagenase perfusion of the liver, was
published. The original method described by Berry and Friend has
undergone many minor modifications by other workers, and the
two-step procedure introduced by Seglen in 1976 has become the most
frequent way to prepare hepatocyte suspensions. An important
development introduced by Bissell in 1973 was the use of the cells
as the first step in monolayer culture. The availability of the
isolated hepatocyte preparation as cells in suspension or culture
has undoubtedly facilitated research on the liver. This was
emphasised in our book, published (with Dr. Greg Barritt) in 1990,
which described in detail methods of preparation and the properties
of the isolated hepatocytes. It also discussed the usefulness of
the preparation for the study of intermediary and xenobiotic
metabolism, calcium ion transport, and the growth and
differentiation of hepatocytes in culture. The book also touched
briefly on a range of specialised techniques, including peri
fusion, subcellular fractionation, transplantation,
cryopreservation and measurement of intracellular pH. Although
standard procedures for the manipulation of hepatocytes have not
changed a great deal in ten years, they have undoubtedly been
refined. This applies particularly to hepatocyte culture
techniques, cryopreservation, and even to preparation of hepatocyte
suspensions, where it is now feasible to use purified enzymes.
There is also much more emphasis on the use and study of human
hepatocytes, particularly in the field of pharmacology and
therapeutics.
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