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The importance of proline on long‐term hepatocyte function in a collagen gel sandwich configuration: Regulation of protein secretion
Author(s) -
Lee Jaegwan,
Morgan Jeffrey R.,
Tompkins Ronald G.,
Yarmush Martin L.
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260400214
Subject(s) - hepatocyte , transferrin , secretion , albumin , proline , chemistry , primary culture , in vitro , function (biology) , medicine , biochemistry , microbiology and biotechnology , endocrinology , biology , amino acid
A serum‐free culture system for primary hepatocytes which maintains stabel high‐level hepatocyte function for prolonged periods in culture has been developed. Isolated rat primary hepatocytes were cultured in serum‐free media between two layer of gelled collagen in a sandwich configuration which reinstates the cellular polarity necessary for long‐term function in vitro. Thsee serum‐free hepatocyte cultures maintained near physiological rates of albumin and transferrin secretion for a minimum of 26 days in culture. L ‐Proline was shown to be critical for both the approach to steady state and maximal level of protein secretion. Analysis of does–response data gave K m values of 2.9 and 1.7 μg/mL for albumin and transferrin secretion, respectively.