A modified method for isolation of bladder cancer stem cells from a MB49 murine cell line.
A modified method for isolation of bladder cancer stem cells from a MB49 murine cell line.
复制标题
从 MB49 小鼠细胞系中分离膀胱癌干细胞的改良方法
DOI：
10.1186/1471-2490-13-57
复制
发表时间：
2013-11-04
期刊：
影响因子：
2
通讯作者：
Tan WL
中科院分区：
文献类型：
--
作者：
Zhu YT;Lei CY;Luo Y;Liu N;He CW;Chen W;Li F;Deng YJ;Tan WL
背景在早期的研究中，该疫苗对膀胱癌有效。然而，一段时间后，由于癌症干细胞无法消除，部分小鼠膀胱肿瘤由于消退而重新生长。方法通过比较不同无血清培养液（SFM），采用有限稀释法和最佳SFM法相结合的方法分离MB 49膀胱癌干细胞（MCSCs）。通过荧光激活细胞分选、定量聚合酶链反应、western blotting、细胞增殖实验、软琼脂实验、transwell实验、化疗抗性实验和肿瘤异种移植实验对MCSCs进行鉴定（20 ng/ml）、碱性成纤维细胞生长因子（20 ng/ml）、白血病抑制因子（20 ng/ml）、B-27无血清补充剂（20 μl/ml）和牛血清白蛋白（4 μg/ml）。MCSC具有高表达肿瘤干细胞标志物（CD 133、CD 44、OCT 4、NANOG和ABCG 2）和分化能力。在功能比较中，MCSCs具有较高的增殖能力，较低的化疗敏感性，更大的迁移在体外，和更强的致瘤能力在vivo.ConclusionMCSCs显示特定的癌症干细胞特性。我们的研究表明，MCSCs成功地分离与有限稀释和SFM方法相结合的改进方法。
BackgroundThe vaccine was efficiently effective against bladder cancer in earlier studies. However, a part of the mouse bladder tumour regrew due to regression after a period of time as the cancer stem cells could not be eliminated. In this study, we showed a modified method for the isolation of MB49 bladder cancer stem cells (MCSCs).MethodsThrough a comparison of different serum-free culture mediums (SFM), MCSCs were isolated by a combination of the limited dilution method and the optimal SFM method. The characterizations of MCSCs were verified by the fluorescence activated cell sorting, the quantitative polymerase chain reaction, the western blotting, the cell proliferation assay, the soft agar assay, the transwell assay, the resistance to chemotherapy assay and the tumor xenograft formation assay.ResultsThe optimal SFM contained a RPMI1640+ epidermal growth factor (20 ng/ml), a basic fibroblast growth factor (20 ng/ml), a leukemia inhibitory factor (20 ng/ml), a B-27 serum-free supplement (20 μl/ml), and a bovine serum albumin (4 μg/ml). MCSCs possessed the high expression of cancer stem cell markers (CD133, CD44, OCT4, NANOG, and ABCG2) and the ability of differentiation. In functional comparisons, MCSCs had higher proliferative abilities, lower susceptibility to chemotherapy, greater migration in vitro, and stronger tumorigenic abilities in vivo.ConclusionMCSCs displayed specific cancer stem cells properties. Our study showed MCSCs were isolated successfully with a modified method using a combination of limited dilution and SFM methods.
登录
查看更多内容
DOI：
10.1007/s11596-012-0043-5
发表时间：
2012-04-01
影响因子：
--
作者：
Gong, Chen;Liao, Hui;Qi, Jun
通讯作者：
Qi, Jun
影响因子：
4.3
作者：
Jung S;Panchalingam KM;Rosenberg L;Behie LA
通讯作者：
Behie LA
HOXB7 作为结直肠癌进展的预后因素和调节因子
影响因子：
11.5
作者：
Liao, Wen-Ting;Jiang, Dan;Ding, Yan-Qing
通讯作者：
Ding, Yan-Qing
DOI：
10.1038/nrurol.2012.142
发表时间：
2012-10
期刊：
Nature reviews. Urology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1016/j.bbrc.2008.04.038
发表时间：
2008-06-20
影响因子：
3.1
作者：
Sung, Ji-Min;Cho, Hee-Jung;Shin, Ho-Chul
通讯作者：
Shin, Ho-Chul