EFFECTIVE CRYOPRESERVATION AND LONG-TERM STORAGE OF PRIMARY HUMAN HEPATOCYTES WITH RECOVERY OF VIABILITY, DIFFERENTIATION, AND REPLICATIVE POTENTIAL
EFFECTIVE CRYOPRESERVATION AND LONG-TERM STORAGE OF PRIMARY HUMAN HEPATOCYTES WITH RECOVERY OF VIABILITY, DIFFERENTIATION, AND REPLICATIVE POTENTIAL
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DOI：
10.1016/0963-6897(95)02001-2
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发表时间：
1995-11-01
影响因子：
3.3
通讯作者：
LEDLEY, FD
中科院分区：
文献类型：
--
作者：
ADAMS, RM;WANG, M;LEDLEY, FD
尽管有成功冷冻保存原代人肝细胞的报道，但现有方法无法产生足够的活细胞回收以满足肝细胞移植基础研究或临床试验的需要。我们现在描述了一种使用威斯康星大学 (UW) 溶液、胎牛血清和二甲基亚砜 (DMSO) 有效冷冻保存原代人肝细胞的方案，该方法在冷冻保存后 8 个月提供 >90% 的分化原代人肝细胞活力（通过台盼蓝排除法测量），保留肝细胞形态、肝脏特异性基因表达（cu、抗胰蛋白酶）和复制。 UW 溶液作为冷冻保存剂的有效性表明，代谢和超微结构因素对于原代人肝细胞的有效冷冻保存可能很重要。本方法代表了冷冻保存分化的原代人肝细胞用于研究的有效方案。该方法可以对人类肝细胞进行表征和储存，用于临床应用，包括肝细胞移植和肝脏辅助装置。
Despite reports of successful cryopreservation of primary human hepatocytes, existing methods do not produce sufficient recovery of viable cells to meet the needs of basic research or clinical trials of hepatocellular transplantation. We now describe a protocol for efficient cryopreservation of primary human hepatocytes using University of Wisconsin (UW) solution, fetal bovine serum, and dimethyl sulfoxide (DMSO), This method provides >90% viability of differentiated, primary human hepatocytes 8 mo after cryopreservation as measured by trypan blue exclusion, preserves hepatocyte morphology, liver-specific gene expression (cu, antitrypsin), and replication. The effectiveness of UW solution as a cryopreservative agent suggests that metabolic as well as ultrastructural factors may be important in the effective cryopreservation of primary human hepatocytes. The present method represents an effective protocol for cryopreserving differentiated primary human hepatocytes for research. This method may allow characterization and banking of human hepatocytes for clinical applications, including hepatocellular transplantation and hepatic assist devices.