Three key variables involved in feeder preparation for the maintenance of human embryonic stem cells

被引：15
作者：
Zhou, Di [1 ]
Liu, Tiancheng [1 ]
Zhou, Xiaoying [1 ]
Lu, Guangxiu [1 ]
机构：
[1] Cent S Univ, Inst Reprod & Stem Cell Engn, Natl Engn & Res Ctr Human Stem Cells, Changsha 410078, Hunan, Peoples R China
来源：
CELL BIOLOGY INTERNATIONAL | 2009年 / 33卷 / 07期
关键词：
Embryonic stem cell; Mouse embryonic fibroblast; Mitomycin C; IN-VITRO; HUMAN BLASTOCYSTS; MOUSE EMBRYOS; CULTURE; LINES; DIFFERENTIATION; MITOMYCIN;
D O I：
10.1016/j.cellbi.2009.04.008
中图分类号：
Q2 [细胞生物学];
学科分类号：
071009 ; 090102 ;
摘要：
Although the development of a feeder-free culture system for future applications of human embryonic stem cells (hESCs), at present the regular culture system uses mitotically inactivated mouse embryonic fibroblasts (mEFs) as feeder cells for maintaining undifferentiated hESCs. Mitomycin C (MMC) is used to inactivate mEFs, but this causes DNA damage, and it is unclear whether MMC remains in the culture system after several washes. Three variables have been evaluated with respect to feeder preparation and MMC involvement, including mEF exposure to MMC, density of feeder cells, and different wash steps during the preparation of feeder cells. These variables are critical to the subsequent planting of hESCs because remnants of MMC would be unsafe with respect to long-term culture of hESCs The novel data here evaluates the remnant amounts of MMC in a hESCs culture system using HPLC/MS/MS. The ultimate objective of this study is the control of MMC within a safe range. (c) 2009 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码：796 / 800
页数：5
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