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Expression of c-kit receptor in human cholangiocarcinoma and in vivo treatment with imatinib mesilate in chimeric mice

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World Journal of Gastroenterology 2006年第10期12卷 1583-1590页

作者： Thomas Kamenz Karel Caca Thilo Blüthner Andrea Tannapfel Joachim Mssner Marcus Wiedmann Department of Internal Medicine Ⅱ University of Leipzig Philipp-Rosenthal-Str.2704103 LeipzigGermany Institute of Pathology University of Leipzig Liebigstr.2604103 LeipzigGermany

AIM： To investigate the c-kit expression in biliary tract cancer cell lines and histological sections from patients with extrahepatic cholangiocarcinoma （CC） and to evaluate the efficacy of in vitro and in vitro treatment with imatinib mesilate. METHODS： The protein expression of c-kit in the human biliary tract cancer cell lines Mz-ChA-2 and EGI-1 and histological sections from 19 patients with extrahepatic CC was assessed by immunoblotting, immunocytochemistry, and immunohistochemistry. The anti-proliferative effect of imatinib mesilate on biliary tract cancer cell lines Mz-ChA-2 and EGI-1 was studied in vitro by automated cell counting. In addition, immunodeficient NMRI mice （Taconic^TM） were subcutaneously injected with 5 × 10^6 cells of cell lines MzChA-2 and EGI-1. After having reached a tumour volume of 200 mm^3, daily treatment was started intraperitoneally with imatinib mesilate at a dose of 50 mgikg or normal saline （NS）. Tumor volume was calculated with a Vernier caliper. After 14 d, mice were sacrificed with tumors excised and tumor mass determined.RESULTS： Immunoblotting revealed presence of c-kit in Mz-ChA-2 and absence in EGI-1 cells. Immunocytochemistry with c-kit antibodies displayed a cytoplasmatic and membraneous localization of receptor protein in Mz-ChA-2 cells and absence of c-kit in EGI-1 cells, c-kit was expressed in 7 of 19 （37%） extrahepatic humanCC tissue samples, 2 showed a moderate and 5 a rather weak immunostaining. Imatinib mesilate at a low concentration of 5 μmoliL caused a significant growth inhibition in the c-kit positive cell line Mz-ChA-2 （31%）, but not in the c-kit negative cell line EGI-1 （0%）（P〈0.05）. Imatinib mesilate at an intermediate concentration of 10 μmoliL inhibited cellular growth of both cell lines （51% vs 57%）. Imatinib mesilate at a higher concentration of 20 μmoliL seemed to have a general toxic effect on both cell lines. The IC50 values were 9.7 μmoliL and 11 μmoliL, respectively. After 14 d of in vitro treat

关键词： Cholangiocarcinoma Imatinib Tyrosine kinase inhibitor c-kit chimeric mice

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Efficient production of chimeric mice from embryonic stem cells injected into 4-to 8-cell and blastocyst embryos

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Journal of Animal Science and Biotechnology 2013年第4期4卷 264-270页

作者： Minhua Hu Hengxi Wei Jingfeng Zhang Yinshan Bai Fenglei Gao Li Li Shouquan Zhang Agricultural Animal Genomics and Molecular Breeding Key Lab of Guangdong Province College of Animal Science South China Agricultural University

Background： Production of chimeric mice is a useful tool for the elucidation of gene function. After successful isolation of embryonic stem （ES） cell lines, there are many methods for producing chimeras, including co-culture with the embryos, microinjection of the ES ceils into pre-implantation embryos, and use of tetraploid embryos to generate the full ES-derived transgenic mice. Here, we aimed to generate the transgenic ES cell line, compare the production efficiency of chimeric mice and its proportion to yield the male chimeric mice by microinjected ES cells into 4- to 8-cell and blastocysts embryos with the application of Piezo-Micromanipulator （PMM）, and trace the fate of the injected ES cells. Results： We successfully generated a transgenic ES cell line and proved that this cell line still maintained pluripotency. Although we achieved a satisfactory chimeric mice rate, there was no significant difference in the production of chimeric mice using the two different methods, but the proportion of the male chimeric mice in the 4- to 8-cell group was higher than in the blastocyst group. We also found that there was no tendency for ES cells to aggregate into the inner cell mass using in vitro culture of the chimeric embryos, indicating that they aggregated randomly. Conclusions： These results showed that the PMM method is a convenient way to generate chimeric mice and microinjection of ES cells into 4- to 8-cell embryos can increase the chance of yielding male chimeras compared to the blastocyst injection. These results provide useful data in transgenic research mediated by ES cells.

关键词： chimeric mice Embryonic stem cell Microinjection

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正常胚胎接种畸胎癌细胞后所发育的嵌合子小鼠(chimeric mice)

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陕西新医药 1979年第6期 66-66页

作者：司履生

把胚胎癌细胞注入小鼠的囊胚期胚胎,以观察:1.在正常胚胎环境下,胚胎癌细胞是否也会像正常胚胎细胞一样分化;2.注射的胚胎癌细胞所繁衍出来的细胞能否形成一般不见于畸胎癌而正常却见于胚胎的那些成分(如肝、肾、淋巴组织)。

关键词：胚胎癌细胞畸胎癌 chimeric mice 小鼠小家鼠

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Pluripotent stem cells secrete Activin A to improve their epiblast competency after injection into recipient embryos

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Protein & Cell 2018年第8期9卷 717-728页

作者： Jinzhu Xiang Suying Cao Liang Zhong Hanning Wang Yangli Pei Qingqing Wei Bingqiang Wen Haiyuan Mu Shaopeng Zhang Liang Yue Genhua Yue Bing Lim Jianyong Han State Key Laboratory for Agrobiotechnology College of Biological Sciences China Agricultural University Beijing 100193 China Animal Science and Technology College Beijing University of Agriculture Beijing 102206 China Temasek Life Sciences Laboratory National University of Singapore Singapore 117604 Singapore Stem Cell and Developmental Biology Genome Institute of Singapore Singapore 138672 Singapore Beijing Advanced Innovation Center for Food Nutrition and Human Health China Agricultural University Beijing 100193 China State Key Laboratory of Animal Nutrition Institute of Animal Sciences Chinese Academy of Agricultural Sciences Beijing 100193 China

It is not fully clear why there is a higher contribution of pluripotent stem cells （PSCs） to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells （ESCs） but also induced pluripotent stem cells （iPSCs） can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a ＂dose effect＂. Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast （EPI） lineage development while promoting trophectoderm （TE） differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A （cultured from 4-cell stage to early blastocyst at E3.5） could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improvetheir EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understand- ing of PSCs effects on early embryo development.

关键词： pluripotent stem cells 4-cell embryoinjection secreted proteins Activin A chimeric mice

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