The enzymic isolation of adult rat hepatocytes in a functional and viable state

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Various methods for the preparation of rat hepatocyte suspensions have been compared with regard to cell yield and viability index. A modified method of collagenase/hyaluronidase digestion using a sequential removal and replacement of Ca2+ and the omission of all perfusion steps has been evolved, thereby making the technique potentially viable for use with pieces of liver such as isolated human liver biopsy material. These suspensions contain large numbers of viable cells, and the viability index is also high (>70%). A number of metabolic functions can be readily detected in these cell suspensions, indicating the intactness of the isolated cells. A systematic survey of the enzymic components required for liver digestion is also reported.

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    Present address: Institut für Physiol. Chemie (Biochemie II), Hoppe-Seyler-Strasse 1, 74 Tübingen, West Germany.

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