title: Cryopreserved mouse fetal liver stromal cells treated with mitomycin C are able to support the growth of human embryonic stem cells.
abstract: An immortalized mouse fetal liver stromal cell line, named KM3, has demonstrated the potential to support the growth and maintenance of human embryonic stem cells (hESCs). In this study, the characteristics of KM3 cells were examined following cryopreservation at -70°C and in liquid nitrogen for 15, 30 and 60 days following treatment with 10 μg/ml mitomycin C. In addition, whether the KM3 cells were suitable for use as feeder cells to support the growth of hESCs was evaluated. The inhibition of mitosis without cell death was observed when the KM3 cells were treated with 10 μg/ml mitomycin C for 2 h. The morphology of the KM3 cells cryopreserved in liquid nitrogen for 60 days was not markedly changed, and the cell survival rate was 84.60±1.14%. By contrast, the survival rate of the KM3 cells was 66.40±2.88% following cryopreservation at -70°C for 60 days; the cells readily detached, were maintained for a shorter time, and had a reduced expression level of basic fibroblast growth factor. hESCs cultured on KM3 cells cryopreserved in liquid nitrogen for 60 days showed the typical bird's nest structure, with clear boundaries and a differentiation rate of 16.33±2.08%. The differentiation rate of hESCs cultured on KM3 cells cryopreserved at -70°C for 60 days was 37.67±3.51%. These results indicate that the cryopreserved KM3 cells treated with mitomycin C may be directly used in the subculture of hESCs, and the effect is relatively good with -70°C short-term or liquid nitrogen cryopreservation.
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Jiabo
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Clinical Laboratory, Jiangsu Provincial Hospital on Integration of Chinese and Western Medicine, Nanjing, Jiangsu 210028, P.R. China.
authors.forename: Quanhui
authors.initials: Q
authors.lastname: Ma
authors.affiliation: Maternal and Child Care Service Centre of Lianyungang, Lianyungang, Jiangsu 222006, P.R. China.
authors.forename: Sanqiang
authors.initials: S
authors.lastname: Hu
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China ; Department of Clinical Laboratory, Zhenjiang Centre for Disease Prevention and Control, Zhenjiang, Jiangsu 212003, P.R. China.
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Wen
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Yongbin
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Clinical Laboratory, Zhenjiang Centre for Disease Prevention and Control, Zhenjiang, Jiangsu 212003, P.R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Qian
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
authors.forename: Wenrong
authors.initials: W
authors.lastname: Xu
keywords: KM3; cryopreservation; feeder; human embryonic stem cells; recovery