دورية أكاديمية
أعرض في EDS

Machine perfusion enhances hepatocyte isolation yields from ischemic livers.

التفاصيل البيبلوغرافية
العنوان: Machine perfusion enhances hepatocyte isolation yields from ischemic livers.
المؤلفون: Izamis ML; Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States. Electronic address: mizamis@mgh.harvard.edu., Perk S; Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States. Electronic address: perksin@gmail.com., Calhoun C; Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States. Electronic address: Calhoun.ck@gmail.com., Uygun K; Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States. Electronic address: uygun.korkut@mgh.harvard.edu., Yarmush ML; Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA 02114, United States; Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, United States. Electronic address: ireis@sbi.org., Berthiaume F; Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, United States. Electronic address: fberthia@rci.rutgers.edu.
المصدر: Cryobiology [Cryobiology] 2015 Oct; Vol. 71 (2), pp. 244-55. Date of Electronic Publication: 2015 Jul 16.
نوع المنشور: Evaluation Study; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0006252 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2392 (Electronic) Linking ISSN: 00112240 NLM ISO Abbreviation: Cryobiology Subsets: MEDLINE
أسماء مطبوعة: Publication: <2000- > : Amsterdam : Elsevier
Original Publication: San Diego, CA : Academic Press
مواضيع طبية MeSH: Cell Separation/*methods , Hepatocytes/*transplantation , Perfusion/*instrumentation , Warm Ischemia/*methods, Adenosine ; Allopurinol ; Animals ; Cell Hypoxia ; Female ; Glutathione ; Hepatocytes/cytology ; Insulin ; Liver/cytology ; Liver/drug effects ; Organ Preservation Solutions ; Perfusion/methods ; Raffinose ; Rats ; Rats, Inbred Lew ; Tissue Donors
مستخلص: Background: High-quality human hepatocytes form the basis of drug safety and efficacy tests, cell-based therapies, and bridge-to-transplantation devices. Presently the only supply of cells derives from an inadequate pool of suboptimal disqualified donor livers. Here we evaluated whether machine perfusion could ameliorate ischemic injury that many of these livers experience prior to hepatocyte isolation.
Methods: Non-heparinized female Lewis rat livers were exposed to an hour of warm ischemia (34°C) and then perfused for 3h. Five different perfusion conditions that utilized the cell isolation apparatus were investigated, namely: (1) modified Williams Medium E and (2) Lifor, both with active oxygenation (95%O(2)/5%CO(2)), as well as (3) Lifor passively oxygenated with ambient air (21%O(2)/0.04%CO(2)), all at ambient temperatures (20 ± 2°C). At hypothermic temperatures (5 ± 1°C) and under passive oxygenation were (4) University of Wisconsin solution (UW) and (5) Vasosol. Negative and positive control groups comprised livers that had ischemia (WI) and livers that did not (Fresh) prior to cell isolation, respectively.
Results: Fresh livers yielded 32 ± 9 million cells/g liver while an hour of ischemia reduced the cell yield to 1.6 ± 0.6 million cells/g liver. Oxygenated Williams Medium E and Lifor recovered yields of 39 ± 11 and 31 ± 2.3 million cells/g liver, respectively. The passively oxygenated groups produced 15 ± 7 (Lifor), 13 ± 7 (Vasosol), and 10 ± 6 (UW)million cells/g liver. Oxygenated Williams Medium E was most effective at sustaining pH values, avoiding the accumulation of lactate, minimizing edematous weight gain and producing bile during perfusion.
Conclusions: Machine perfusion results in a dramatic increase in cell yields from livers that have had up to an hour of warm ischemia, but perfusate choice significantly impacts the extent of recovery. Oxygenated Williams Medium E at room temperature is superior to Lifor, UW and Vasosol, largely facilitated by its high oxygen content and low viscosity.
(Copyright © 2015 Elsevier Inc. All rights reserved.)
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معلومات مُعتمدة: R00 DK080942 United States DK NIDDK NIH HHS; R01DK59766 United States DK NIDDK NIH HHS; R01 DK096075 United States DK NIDDK NIH HHS; R00DK088962 United States DK NIDDK NIH HHS; R01 DK059766 United States DK NIDDK NIH HHS; R01DK096075 United States DK NIDDK NIH HHS
فهرسة مساهمة: Keywords: Hepatocytes; Isolation; Lifor; Perfusion; Temperature; UW; Vasosol; Viscosity; Williams E
المشرفين على المادة: 0 (Insulin)
0 (Organ Preservation Solutions)
0 (University of Wisconsin-lactobionate solution)
63CZ7GJN5I (Allopurinol)
GAN16C9B8O (Glutathione)
K72T3FS567 (Adenosine)
N5O3QU595M (Raffinose)
تواريخ الأحداث: Date Created: 20150719 Date Completed: 20160425 Latest Revision: 20231111
رمز التحديث: 20231111
مُعرف محوري في PubMed: PMC4584189
DOI: 10.1016/j.cryobiol.2015.07.006
PMID: 26188080
قاعدة البيانات: MEDLINE
الوصف
تدمد:1090-2392
DOI:10.1016/j.cryobiol.2015.07.006