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Dynamic seeding and in vitro culture of hepatocytes in a flow perfusion system

Cited 112 time in Web of Science Cited 123 time in Scopus
Authors

Stephen S. Kim; Cathryn A. Sundback; Satoshi Kaihara; Mark S. Benvenuto; Byung-Soo Kim; David J. Mooney; Joseph P. Vacanti

Issue Date
2000-02
Publisher
MARY ANN LIEBERT INC PUBL
Citation
TISSUE ENGINEERING, Vol.6 No.1, pp.39-44
Abstract
Our laboratory has investigated hepatocyte transplantation using biodegradable polymer matrices as an alternative treatment to end-stage liver disease. One of the major limitations has been the insufficient survival of an adequate mass of transplanted cells. This study investigates a novel method of dynamic seeding and culture of hepatocytes in a flow perfusion system. In experiment I, hepatocytes were flow-seeded onto PGA scaffolds and cultured in a flow perfusion system for 24 h. Overall metabolic activity and distribution of cells were assessed by their ability to reduce MTT. DNA quantification was used to determine the number of cells attached. Culture medium was analyzed for albumin content. In Experiment II, hepatocyte/polymer constructs were cultured in a perfusion system for 2 and 7 days. The constructs were examined by SEM and histology. Culture medium was analyzed for albumin. In experiment I, an average of 4.4 x 10(6) cells attached to the scaffolds by DNA quantification. Cells maintained a high metabolic activity and secreted albumin at a rate of 13 pg/cell/day. In experiment II, SEM demonstrated successful attachment of hepatocytes on the scaffolds after 2 and 7 days. Cells appeared healthy on histology and maintained a high rate of albumin secretion through day 7. Hepatocytes can be dynamically seeded onto biodegradable polymers and survive with a high rate of albumin synthesis in the flow perfusion culture system.
ISSN
1076-3279
URI
https://hdl.handle.net/10371/204470
DOI
https://doi.org/10.1089/107632700320874
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  • College of Engineering
  • School of Chemical and Biological Engineering
Research Area biomaterials, nanomedicine, regenerative medicine

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