1 Department of Surgery, Campus Charité Mitte | Campus Virchow-Klinikum, Experimental Surgery and Regenerative Medicine, Charité-Universitatsmedizin Berlin , Berlin, Germany .
2 Department of Diagnostic and Interventional Radiology, Campus Virchow-Klinikum, Charité-Universitatsmedizin Berlin , Berlin, Germany .
Tissue Eng Part C Methods. 2018 Mar;24(3):179-186. doi: 10.1089/ten.TEC.2017.0369. Epub 2018 Feb 27.
The need for primary human hepatocytes is constantly growing for basic research, as well as for therapeutic applications. However, the isolation outcome strongly depends on the quality of liver tissue, and we are still lacking a preoperative test that allows the prediction of the hepatocyte isolation outcome. In this study, we evaluated the "maximal liver function capacity test" (LiMAx) as predictive test for the quantitative and qualitative outcome of hepatocyte isolation. This test is already used in clinical routine to measure preoperative and to predict postoperative liver function. The patient's preoperative mean LiMAx was obtained from the patient records, and preoperative computed tomography and magnetic resonance images were used to calculate the whole liver volume to adjust the mean LiMAx. The outcome parameters of the hepatocyte isolation procedures were analyzed in correlation with the adjusted mean LiMAx. Primary human hepatocytes were isolated from partial hepatectomies (n = 64). From these 64 hepatectomies we included 48 to our study and correlated their isolation outcome parameters with volume corrected LiMAx values. From a total of 11 hepatocyte isolation procedures, metabolic parameters (albumin, urea, and aspartate aminotransferase or AST) were assessed during the hepatocyte cultivation period of 5 days. The volume adjusted mean LiMAx showed a significant positive correlation with the total cell yield (p = 0.049; r = 0.242; n = 48). The correlations of volume adjusted LiMAx values with viable cell yield and cell viability did not reach statistical significance. To create a more homogenous study group regarding tumor entities, subgroup analyses were performed. A subgroup analysis of isolations from patients with colorectal metastasis revealed a significant correlation between volume adjusted mean LiMAx and total cell yield (p = 0.012; r = 0.488; n = 21) and viable cell yield (p = 0.034; r = 0.405; n = 21), whereas a subgroup analysis of isolations of patients with carcinoma of the biliary tree showed significant correlations of volume adjusted mean LiMAx with cell viability (r = 0.387; p = 0.046; n = 20) and lacked significant correlations with total cell yield (r = -0.060; p = 0.401; n = 20) and viable cell yield (r = 0.012; p = 0.480; n = 20). The volume-adjusted mean LiMAx did not show a significant correlation with any of the metabolic parameters. In conclusion, the LiMAx test might be a useful tool to predict the quantitative outcome of hepatocyte isolation, as long as underlying liver disease is taken into consideration.
原发性人肝细胞的需求不断增长，无论是用于基础研究还是治疗应用。然而，分离的结果在很大程度上取决于肝组织的质量，我们仍然缺乏一种术前测试，可以预测肝细胞分离的结果。在这项研究中，我们评估了"最大肝功能容量测试"(LiMAx)作为预测肝细胞分离定量和定性结果的测试。该测试已在临床常规中用于测量术前和预测术后肝功能。患者的术前平均 LiMAx 从患者记录中获得，并使用术前计算机断层扫描和磁共振成像来计算整个肝脏体积以调整平均 LiMAx。将肝细胞分离程序的结果参数与调整后的平均 LiMAx 相关联进行分析。从部分肝切除术（n=64）中分离出原发性人肝细胞。我们从这 64 例肝切除术中有 48 例纳入了我们的研究，并将其分离结果参数与体积校正的 LiMAx 值相关联。在总共 11 次肝细胞分离过程中，在 5 天的肝细胞培养期间评估了代谢参数（白蛋白、尿素和天冬氨酸氨基转移酶或 AST）。体积校正的平均 LiMAx 值与总细胞产量呈显著正相关（p=0.049；r=0.242；n=48）。体积校正的 LiMAx 值与有活力的细胞产量和细胞活力之间的相关性没有达到统计学意义。为了创建一个关于肿瘤实体的更同质的研究组，进行了亚组分析。对来自结直肠转移患者的分离物进行亚组分析显示，体积校正的平均 LiMAx 值与总细胞产量（p=0.012；r=0.488；n=21）和有活力的细胞产量（p=0.034；r=0.405；n=21）之间存在显著相关性，而对胆管癌患者分离物进行的亚组分析显示，体积校正的平均 LiMAx 值与细胞活力（r=0.387；p=0.046；n=20）存在显著相关性，而与总细胞产量（r=-0.060；p=0.401；n=20）和有活力的细胞产量（r=0.012；p=0.480；n=20）之间无显著相关性。体积校正的平均 LiMAx 值与任何代谢参数均无显著相关性。总之，只要考虑到潜在的肝病，LiMAx 测试可能是预测肝细胞分离定量结果的有用工具。
