Cryopreserved neonatal hepatocytes may be a source for transplantation: Evaluation of functionality toward clinical use
Cryopreserved neonatal hepatocytes may be a source for transplantation: Evaluation of functionality toward clinical use
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DOI：
10.1002/lt.25015
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发表时间：
2018-03-01
影响因子：
4.6
通讯作者：
Fitzpatrick, Emer
中科院分区：
文献类型：
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作者：
Lee, Charlotte A.;Dhawan, Anil;Fitzpatrick, Emer
新生儿肝脏是临床移植的优质肝细胞的潜在来源。我们比较了新生肝细胞（NH）和成人肝细胞（AH）的活力和功能，并报告了它们在门静脉内和藻酸盐微珠中的临床应用。从供体肝脏分离后，使用白蛋白、α-1-抗胰蛋白酶和因子VII评估肝细胞功能。通过测定试卤灵结合、氨代谢、尿苷二磷酸葡萄糖醛酸转移酶活性和诱导后细胞色素P450（CYP）功能来研究代谢功能。使用体外血液灌注模型研究了NH和AH对即时血液介导的炎症反应的激活，并使用实时聚合酶链反应（RT-PCR）分析了组织因子表达。临床肝细胞移植（HT）采用标准方案进行。从14个新生儿肝脏中分离肝细胞，平均活力为89.4% +/- 1.8%（平均值+/-平均值的标准误差），平均产量为9.3 x 10（6）+/- 2.0 x 10（6）个细胞/g。从14个成人肝脏中分离肝细胞，平均活力为78.6% +/- 2.4%，产量为2.2 x 10（6）+/- 0.5 x 10（5）个细胞/g。NHs在冷冻保存后的存活率显著高于AH，具有更好的附着效率和更少的质膜渗漏。NH和AH之间的白蛋白、α-1-抗胰蛋白酶和因子VII合成无差异（P > 0.05）。新生儿细胞具有可诱导的第1阶段酶，如通过细胞功能和功能性第2阶段酶评估的，其中活性与AH相当。在体外血液灌注模型中，与NH相比，AH引起血栓形成增加，血小板和白色细胞消耗更多（28.3 x 10（9）vs. 118.7 x 10（9）和3.3 x 10（9）vs. 6.6 x 10（9）; P < 0.01）。门静脉内移植和腹腔内移植藻酸盐包封的肝细胞是安全的，初步数据表明，细胞可能激活免疫反应的程度低于成年细胞。总之，我们已经表明NHs具有优异的细胞活力、功能和药物代谢，使其成为临床HT的合适替代来源。肝移植24 394-406 2018 AASLD。
Neonatal livers are a potential source of good-quality hepatocytes for clinical transplantation. We compared viability and function of neonatal hepatocytes (NHs) and adult hepatocytes (AHs) and report their clinical use both intraportally and in alginate microbeads. Following isolation from donor livers, hepatocyte function was assessed using albumin, alpha-1-antitrypsin, and factor VII. Metabolic function was investigated by measuring resorufin conjugation, ammonia metabolism, uridine diphosphate glucuronosyltransferase enzyme activity, and cytochrome P450 (CYP) function following induction. Activation of the instant blood-mediated inflammatory reaction by NHs and AHs was investigated using an in vitro blood perfusion model, and tissue factor expression was analyzed using real-time polymerase chain reaction (RT-PCR). Clinical hepatocyte transplantation (HT) was undertaken using standard protocols. Hepatocytes were isolated from 14 neonatal livers, with an average viability of 89.4% +/- 1.8% (mean +/- standard error of the mean) and average yield of 9.3 x 10(6) +/- 2.0 x 10(6) cells/g. Hepatocytes were isolated from 14 adult livers with an average viability of 78.6% +/- 2.4% and yield 2.2 x 10(6) +/- 0.5 x 10(5)cells/g. NHs had significantly higher viability after cryopreservation than AHs, with better attachment efficiency and less plasma membrane leakage. There were no differences in albumin, alpha-1-antitrypsin, and factor VII synthesis between NHs and AHs (P > 0.05). Neonatal cells had inducible phase 1 enzymes as assessed by CYP function and functional phase 2 enzymes, in which activity was comparable to AHs. In an in vitro blood perfusion model, AHs elicited increased thrombus formation with a greater consumption of platelets and white cells compared with NHs (28.3 x 10(9) versus 118.7 x 10(9) and 3.3 x 10(9) versus 6.6 x 10(9); P < 0.01). Intraportal transplantation and intraperitoneal transplantation of alginate encapsulated hepatocytes was safe, and preliminary data suggest the cells may activate the immune response to a lesser degree than adult cells. In conclusion, we have shown NHs have excellent cell viability, function, and drug metabolism making them a suitable alternative source for clinical HT. Liver Transplantation 24 394-406 2018 AASLD.