| Studies in rat liver perfusion for optimal harvest of hepatocytes. | |
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MedLine Citation:
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PMID: 2153793 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Pediatric liver transplantation is successful but donor scarcity is a major limitation. We are studying hepatocyte transplantation as an alternative to provide functional hepatic replacement. This report details the study of rat liver perfusion for optimal harvest of hepatocytes and cell implantation. We performed 128 rat liver perfusions using a technique modified from the two-step enzymatic perfusion described by Seglen. We examined variations in the perfusion, rate, time, antegrade versus retrograde, pulsatile versus continuous flow, temperature, collagenase type, and variables of buffer composition. We have found optimal cell yield and viability under the following conditions: in situ perfusion, continuous flow at 25 cc/min, retrograde perfusion via the inferior vena cava, water bath temperature 38 degrees C, Boerhinger-Mannheim collagenase using a nonoxygenated HEPES based perfusion buffer, pH 7.4, for the initial perfusion and the same buffer with 4.8 mmol/L CaCl2 for the collagenase perfusion. These conditions consistently generate cell harvests of 500 to 700 x 10(5) cells/g of liver tissue with cell viability between 85% and 95%. |
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Authors:
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J Aiken; L Cima; B Schloo; D Mooney; L Johnson; R Langer; J P Vacanti |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of pediatric surgery Volume: 25 ISSN: 0022-3468 ISO Abbreviation: J. Pediatr. Surg. Publication Date: 1990 Jan |
Date Detail:
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Created Date: 1990-03-15 Completed Date: 1990-03-15 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0052631 Medline TA: J Pediatr Surg Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 140-4; discussion 144-5 Citation Subset: IM |
Affiliation:
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Department of Surgery, Children's Hospital, Boston, MA 02115. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Separation / methods* Cells, Cultured Liver / cytology* Liver Diseases / therapy Liver Transplantation* Microbial Collagenase / administration & dosage Perfusion / methods* Rats Rats, Inbred F344 Rats, Inbred Strains |
| Chemical | |
Reg. No./Substance:
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EC 3.4.24.3/Microbial Collagenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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