一种改进的、高细胞活力和低细胞损失的低温保存肝细胞的纯化方法。

An improved purification approach with high cell viability and low cell loss for cryopreserved hepatocytes.

机构信息

Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

Cryobiology. 2010 Apr;60(2):238-9. doi: 10.1016/j.cryobiol.2009.11.003. Epub 2009 Nov 20.

DOI:10.1016/j.cryobiol.2009.11.003

PMID:19932092

Abstract

A modified purification procedure is described for effectively eliminating dead cells after hepatocyte cryopreservation. Isolated hepatocytes from six pig tissue samples were cryopreserved in liquid nitrogen for 2 weeks. After thawing, we developed a pre-incubation step prior to gradient centrifugation. The hepatocytes were subsequent cultured in suspension overnight (12-16 h), and then dead cells were eliminated by Ficoll 400 purification. The results showed that a high viability (mean of 96%) of cells was obtained, with a low viable cell loss in number (2-5%), by using this modified method.

摘要

本文描述了一种改良的纯化程序，可在肝细胞冷冻保存后有效去除死亡细胞。从六个猪组织样本中分离的肝细胞在液氮中冷冻保存 2 周。解冻后，我们在梯度离心前开发了一个预孵育步骤。随后将肝细胞在悬浮液中培养过夜（12-16 小时），然后用 Ficoll 400 纯化去除死细胞。结果表明，使用这种改良方法可获得高存活率（平均值为 96%），活细胞数量损失低（2-5%）。