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Effect of Down-regulated Expression of cPouV and cNanog on Pluripotency of Chicken(Gallus gallus) Embryonic Stem Cells(cESCs)
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作者 zhisheng chen Te PENG +4 位作者 Shengfeng chen Dongsheng LI Huiqin JI Bingyun WANG Jinding chen 《Agricultural Biotechnology》 CAS 2015年第3期53-58,共6页
To explore the pluripotency maintenance and update the functional influence of pluripotency genes cNanog and cPouV in chicken ( C,a/lus gallus) embry- onic stem cells ( cESCs), the stable RNAi vectors pSuper-cNano... To explore the pluripotency maintenance and update the functional influence of pluripotency genes cNanog and cPouV in chicken ( C,a/lus gallus) embry- onic stem cells ( cESCs), the stable RNAi vectors pSuper-cNanog and pSuper-cPouV constructed previously were used to transfect cESCs. The mRNA levels of two target genes were detected with real- time PCR. These two genes were down-regulated since the 48^th and the down-reg-lation continued with the extension of time, the interference efficiency reached 65% at 96^th hour (P 〈0.05). With the down-regulation of cNanog or cPouV gene, cESCs showed differentiation and prolifera- tion rate of these cells slowed down, the domed colony of these cells disappeared gradually when the edge of colony became irregular. At 96^th hour after transfection, the alkline phosphatase (AKP) and stage-specific embryonic antigen-1 ( SSEA-1 ) were not be detected in cNanog gene-knecked out eESCs, but it was done in that with cPouV gene -knocked out. The cPouV-suppressing cESCs were again transfected with pSuper-cNanog, the pluripotency markers AKP and SSEA-1 were both not found expressing at the 48^th hour. The results showed that cPouV and cNartog genes played an important role in maintaining pluripotency and self- renewal in cESCs, and cNanog gene was dominant. To sum up, our results may provide insights into the molecular regulation mechanism of avian during development. 展开更多
关键词 Chicken embryonic stem cells cNanog cPouV PLURIPOTENCY RNA interference
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Chicken Mesenchymal Stem Cells as Feeder Cells Facilitate the Cultivation of Primordial Germ Cells from Circulating Blood and Gonadal Ridge
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作者 Dongsheng Li zhisheng chen +5 位作者 Shengfeng chen Huiqin Ji Xiaoshu Zhan Dongzhang Luo Huina Luo Bingyun Wang 《Stem Cell Discovery》 2019年第1期1-14,共14页
Long-term maintenance of chicken primordial germ cells (PGCs) in vitro has tremendous potential for transgenic chicken production. Feeder cells are essential for the establishment and culture of chicken PGCs in vitro.... Long-term maintenance of chicken primordial germ cells (PGCs) in vitro has tremendous potential for transgenic chicken production. Feeder cells are essential for the establishment and culture of chicken PGCs in vitro. Buffalo rat liver (BRL) cells are the most commonly used feeder cells for PGCs culture;however, this feeder layers from other animal species usually cause immunogenic contaminations, compromising the potential of PGCs in applications. Therefore, we tested chicken source mensenchymal stem cell (MSCs) derived from bone marrow as feeder cells to further improve PGC culture conditions. MSCs derived from chicken bone marrow have a powerful capacity to proliferate and secrete cytokines. We found chicken primordial germ cells derived from circulating blood (cPGCs) and gonads (gPGCs) can be maintained and proliferated with MSCs feeder layer cells. PGCs co-cultured on MSCs feeder retained their pluripotency, expressed PGCs specific genes and stemness markers, and maintained undifferentiated state. Our study indicated that the xeno-free MSCs-feeders culture system is a good candidate for growth and expansion of PGCs as the stepping stone for transgenic chicken research. 展开更多
关键词 CHICKEN Primordial GERM CELLS Mensenchymal Stem CELLS TRANSGENIC
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耻脐入路单孔腹腔镜后鞘后技术在脐疝修补术中的应用
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作者 陈志升 司仙科 +7 位作者 吴卫东 李森 杨佳华 李炜 郑侃 于昆 罗晓睿 葛卫红 《中华疝和腹壁外科杂志(电子版)》 2024年第2期184-188,共5页
目的探讨耻脐入路单孔腹腔镜后鞘后修补术治疗脐疝的临床应用价值。方法回顾性研究2021年6月至2023年8月,上海中医药大学附属普陀医院微创外科收治的18例实施耻脐入路单孔腹腔镜后鞘后疝修补术的脐疝患者,根据临床病例资料、手术操作的... 目的探讨耻脐入路单孔腹腔镜后鞘后修补术治疗脐疝的临床应用价值。方法回顾性研究2021年6月至2023年8月,上海中医药大学附属普陀医院微创外科收治的18例实施耻脐入路单孔腹腔镜后鞘后疝修补术的脐疝患者,根据临床病例资料、手术操作的技术细节及术后随访情况,分析该术式的临床效果及获益。结果18例患者均顺利完成手术,无中转开放手术,平均手术时间(106.67±28.49)min,术后6~8 h下床活动,术后8、24 h视觉模拟评分分别为2.94、2.50分,术后平均住院时间为(2.02±0.63)d;术后均无出血、肠梗阻、肠瘘、脐部坏死等严重并发症;随访时间为3~28个月,随访率100%,无复发。结论耻脐入路行单孔腹腔镜下后鞘后脐疝修补术安全可行,隐瘢痕效果好,术后疼痛轻,恢复快,可作为腹腔镜下脐疝修补的一种选择术式。 展开更多
关键词 脐疝 耻脐入路 腹腔镜 单孔 后鞘后间隙 疝修补术
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Quantitative proteomics analysis of parthenogenetically induced pluripotent stem cells
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作者 Zhe Hu LeiWang +9 位作者 Zhensheng Xie Xinlei Zhang Du Feng Fang Wang Bingfeng Zuo Lingling Wang Zhong Liu zhisheng chen Fuquan Yang Lin Liu 《Protein & Cell》 SCIE CSCD 2011年第8期631-646,共16页
Parthenogenetic embryonic stem(pES)cells isolated from parthenogenetic activation of oocytes and embryos,also called parthenogenetically induced pluripotent stem cells,exhibit pluripotency evidenced by both in vitro a... Parthenogenetic embryonic stem(pES)cells isolated from parthenogenetic activation of oocytes and embryos,also called parthenogenetically induced pluripotent stem cells,exhibit pluripotency evidenced by both in vitro and in vivo differentiation potential.Differential proteomic analysis was performed using differential in-gel electrophoresis and isotope-coded affinity tag-based quantitative proteomics to investigate the molecular mechanisms underlying the developmental pluripotency of pES cells and to compare the protein expression of pES cells generated from either the in vivo-matured ovulated(IVO)oocytes or from the in vitro-matured(IVM)oocytes with that of fertilized embryonic stem(fES)cells derived from fertilized embryos.A total of 76 proteins were upregulated and 16 proteins were downregulated in the IVM pES cells,whereas 91 proteins were upregulated and 9 were downregulated in the IVO pES cells based on a minimal 1.5-fold change as the cutoff value.No distinct pathways were found in the differentially expressed proteins except for those involved in metabolism and physiological processes.Notably,no differences were found in the protein expression of imprinted genes between the pES and fES cells,suggesting that genomic imprinting can be corrected in the pES cells at least at the early passages.The germline competent IVM pES cells may be applicable for germ cell renewal in aging ovaries if oocytes are retrieved at a younger age. 展开更多
关键词 parthenogenetic embryonic stem cell PROTEOME fluorescent two-dimensional difference in-gel electrophoresis isotope-coded affinity tag
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