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Human Hepatocyte Isolation: Does Portal Vein Embolization Affect the Outcome?

Percoll公司 胶原酶 灌注 肝细胞 白蛋白 医学 栓塞 肝细胞生长因子 肝再生 内科学 病理 生物 泌尿科 外科 再生(生物学) 生物化学 离心 体外 受体 细胞生物学
作者
Martin Kluge,Anja Reutzel‐Selke,Hendrik Napierala,Karl H. Hillebrandt,Rebeka Dalma Major,Benjamin Struecker,Annekatrin Leder,J. Siefert,Peter Tang,Steffen Lippert,Hannes Sallmon,Daniel Seehofer,Johann Pratschke,Igor M. Sauer,Nathanael Raschzok
出处
期刊:Tissue Engineering Part C-methods [Mary Ann Liebert, Inc.]
卷期号:22 (1): 38-48 被引量:9
标识
DOI:10.1089/ten.tec.2015.0190
摘要

Primary human hepatocytes are widely used for basic research, pharmaceutical testing, and therapeutic concepts in regenerative medicine. Human hepatocytes can be isolated from resected liver tissue. Preoperative portal vein embolization (PVE) is increasingly used to decrease the risk of delayed postoperative liver regeneration by induction of selective hypertrophy of the future remnant liver tissue. The aim of this study was to investigate the effect of PVE on the outcome of hepatocyte isolation. Primary human hepatocytes were isolated from liver tissue obtained from partial hepatectomies (n = 190) using the two-step collagenase perfusion technique followed by Percoll purification. Of these hepatectomies, 27 isolations (14.2%) were performed using liver tissue obtained from patients undergoing PVE before surgery. All isolations were characterized using parameters that had been described in the literature as relevant for the outcome of hepatocyte isolation. The isolation outcomes of the PVE and the non-PVE groups were then compared before and after Percoll purification. Metabolic parameters (transaminases, urea, albumin, and vascular endothelial growth factor secretion) were measured in the supernatant of cultured hepatocytes for more than 6 days (PVE: n = 4 and non-PVE: n = 3). The PVE and non-PVE groups were similar in regard to donor parameters (sex, age, and indication for surgery), isolation parameters (liver weight and cold ischemia time), and the quality of the liver tissue. The mean initial viable cell yield did not differ between the PVE and non-PVE groups (10.16 ± 2.03 × 10(6) cells/g vs. 9.70 ± 0.73 × 10(6) cells/g, p = 0.499). The initial viability was slightly better in the PVE group (77.8% ± 2.03% vs. 74.4% ± 1.06%). The mean viable cell yield (p = 0.819) and the mean viability (p = 0.141) after Percoll purification did not differ between the groups. PVE had no effect on enzyme leakage and metabolic activity of cultured hepatocytes. Although PVE leads to drastic metabolic alterations and changes in hepatic blood flow, embolized liver tissue is a suitable source for the isolation of primary human hepatocytes and is equivalent to untreated liver tissue in regard to cell yield and viability.
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