AIM： To examine the human hepatic parenchymal and stromal components in rat liver and the phenotypic changes of human cells in liver of human-rat chimera （HRC） generated by in utero transplantation of human cells d...AIM： To examine the human hepatic parenchymal and stromal components in rat liver and the phenotypic changes of human cells in liver of human-rat chimera （HRC） generated by in utero transplantation of human cells during partial hepatectomy （PHx）-induced liver regeneration. METHODS： Human hepatic parenchymal and stromal components and phenotypic changes of human cells during liver regeneration were examined by flow oytometry, in situ hybridization and immunohistochemistry. RESULTS： ISH analysis positive cells in hepatic demonstrated human Aluparenchyma and stroma of recipient liver. Functional human hepatocytes generated in this model potentially constituted human hepatic functional units with the presence of donor-derived human endothelial and biliary duct cells in host liver. Alpha fetoprotein （AFP）^＋, CD34^＋ and CD45^＋ cells were observed in the chimeric liver on day 10 after PHxinduced liver regeneration and then disappeared in PHx group, but not in non-PHx group, suggesting that dynamic phenotypic changes of human cells expressing AFP, CD34 and CD45 cells may occur during the chimeric liver regeneration. Additionally, immunostaining for human proliferating cell nuclear antigen （PCNA） showed that the number of PCNA-positive cells in the chimeric liver of PHx group was markedly increased, as compared to that of control group, indicating that donor-derived human cells are actively proliferated during PHx-induced regeneration of HRC liver. CONCLUSION： HRC liver provides a tool for investigating human liver regeneration in a humanized animal model.展开更多
背景:研究发现人脐血富血小板血浆与人脐血间充质干细胞对薄型子宫内膜有一定治疗效果,但目前针对人脐血单个核细胞对薄型子宫内膜的研究尚未报道,且目前尚缺乏三者比较的相关研究。目的:探讨人脐血富血小板血浆、单个核细胞及间充质干...背景:研究发现人脐血富血小板血浆与人脐血间充质干细胞对薄型子宫内膜有一定治疗效果,但目前针对人脐血单个核细胞对薄型子宫内膜的研究尚未报道,且目前尚缺乏三者比较的相关研究。目的:探讨人脐血富血小板血浆、单个核细胞及间充质干细胞修复大鼠薄型子宫内膜的效果与机制。方法:60只雌性SPF级SD大鼠随机分为假手术组、模型组、人脐血富血小板血浆组、人脐血单个核细胞组和人脐血间充质干细胞组,每组12只。假手术组宫角注射0.5 mL生理盐水,5 min后再灌注0.5 mL PBS;模型组、人脐血富血小板血浆组、人脐血单个核细胞组和人脐血间充质干细胞组宫角注射0.5 mL体积分数95%乙醇,5 min后吸出剩余乙醇,经生理盐水冲洗2次后再分别灌注0.5 mL PBS、人脐血富血小板血浆、人脐血单个核细胞悬液(1×10^(7)个)和人脐血间充质干细胞悬液(1×10^(7)个),于再灌注后第3个正常动情周期的动情间期收集脏器组织与血清,检测相关指标。结果与结论:①子宫组织大体观、苏木精-伊红染色和Masson染色结果:假手术组结构完整,内膜厚度适中,血管结构清晰。与假手术组相比,模型组子宫萎缩,结构不完整,内膜厚度、腺体数量明显减少,血管结构紊乱,纤维化增加;与模型组相比,经人脐血衍生物治疗后,子宫大小、结构、内膜厚度均有所恢复(均P<0.01),并且纤维化均减轻,其中以人脐血单个核细胞组恢复最为明显。在腺体数量增加方面,以人脐血富血小板血浆组效果最为显著(P<0.0001)。②子宫组织免疫组化与免疫荧光结果显示,与假手术组相比,模型组细胞增殖相关指标角蛋白9、波形蛋白,内膜容受性相关指标白血病抑制因子、整合素αγβ3,血小板内皮细胞黏附分子、碱性成纤维细胞生长因子、血管内皮生长因子表达均降低(均P<0.05)。与模型组相比,经人脐血衍生物治疗后以上指标均有显著性升高,人脐血衍生物各组比较:角蛋白9、血管内皮生长因子蛋白:人脐血单个核细胞组>人脐血间充质干细胞组>人脐血富血小板血浆组;波形蛋白、白血病抑制因子蛋白:人脐血间充质干细胞组>人脐血单个核细胞组>人脐血富血小板血浆组;整合素αγβ3蛋白:人脐血富血小板血浆组>人脐血间充质干细胞组>人脐血单个核细胞组;血小板内皮细胞黏附分子蛋白:人脐血富血小板血浆组>人脐血单个核细胞组>人脐血间充质干细胞组;碱性成纤维细胞生长因子蛋白:人脐血单个核细胞组>人脐血富血小板血浆组>人脐血间充质干细胞组。③Western blot检测显示,与假手术组相比,模型组白细胞介素6、白细胞介素1β和肿瘤坏死因子α蛋白表达水平显著升高(均P<0.001),经人脐血衍生物治疗后上述指标均降低(均P<0.05)。④ELISA检测显示,与假手术组相比,模型组抗缪勒氏管激素、雌二醇、孕酮水平降低,促卵泡生成素、促黄体生成素水平增加(除促黄体生成素外,均P<0.05)。经人脐血衍生物治疗后,性激素和抗缪勒氏管激素水平有一定程度恢复。⑤生育力实验显示,与假手术组相比,模型组受孕时间有所增加,产仔数明显降低(均P<0.05)。经人脐血衍生物治疗后,3组产仔数均有所增加(P<0.05),组间比较尚未发现明显差异。此研究初步表明人脐血单个核细胞对薄型子宫内膜有一定治疗效果,且人脐血富血小板血浆、人脐血单个核细胞和人脐血间充质干细胞分别在改善薄型子宫内膜再生功能、内膜容受性、血管生成、炎症调控及妊娠结局方面具有不同的优势与差异。展开更多
基金Supported by The National Natural Science Foundation of China, No. 30271177 and No. 39870676the Major Scienceand Technology Projects of Guangdong Province, No. B602+4 种基金the Natural Science Foundation of Guangdong Province, No.021903the Science and Technology Planning Project of Guangdong Province, No. 2009B060300008the Science and Technology Projects of Guangzhou City, No. 2002Z2E0121the Medical Scientific Research Foundation of Guangdong Province, No. A2007359the Science and Technology Talented Man Foundation of Outstanding Young and Middle-aged People of Southern Medical University,the Special Fund of Scientific Instrument Collaborative Share-net in Guangzhou, No. 2006176
文摘AIM： To examine the human hepatic parenchymal and stromal components in rat liver and the phenotypic changes of human cells in liver of human-rat chimera （HRC） generated by in utero transplantation of human cells during partial hepatectomy （PHx）-induced liver regeneration. METHODS： Human hepatic parenchymal and stromal components and phenotypic changes of human cells during liver regeneration were examined by flow oytometry, in situ hybridization and immunohistochemistry. RESULTS： ISH analysis positive cells in hepatic demonstrated human Aluparenchyma and stroma of recipient liver. Functional human hepatocytes generated in this model potentially constituted human hepatic functional units with the presence of donor-derived human endothelial and biliary duct cells in host liver. Alpha fetoprotein （AFP）^＋, CD34^＋ and CD45^＋ cells were observed in the chimeric liver on day 10 after PHxinduced liver regeneration and then disappeared in PHx group, but not in non-PHx group, suggesting that dynamic phenotypic changes of human cells expressing AFP, CD34 and CD45 cells may occur during the chimeric liver regeneration. Additionally, immunostaining for human proliferating cell nuclear antigen （PCNA） showed that the number of PCNA-positive cells in the chimeric liver of PHx group was markedly increased, as compared to that of control group, indicating that donor-derived human cells are actively proliferated during PHx-induced regeneration of HRC liver. CONCLUSION： HRC liver provides a tool for investigating human liver regeneration in a humanized animal model.
文摘背景:研究发现人脐血富血小板血浆与人脐血间充质干细胞对薄型子宫内膜有一定治疗效果,但目前针对人脐血单个核细胞对薄型子宫内膜的研究尚未报道,且目前尚缺乏三者比较的相关研究。目的:探讨人脐血富血小板血浆、单个核细胞及间充质干细胞修复大鼠薄型子宫内膜的效果与机制。方法:60只雌性SPF级SD大鼠随机分为假手术组、模型组、人脐血富血小板血浆组、人脐血单个核细胞组和人脐血间充质干细胞组,每组12只。假手术组宫角注射0.5 mL生理盐水,5 min后再灌注0.5 mL PBS;模型组、人脐血富血小板血浆组、人脐血单个核细胞组和人脐血间充质干细胞组宫角注射0.5 mL体积分数95%乙醇,5 min后吸出剩余乙醇,经生理盐水冲洗2次后再分别灌注0.5 mL PBS、人脐血富血小板血浆、人脐血单个核细胞悬液(1×10^(7)个)和人脐血间充质干细胞悬液(1×10^(7)个),于再灌注后第3个正常动情周期的动情间期收集脏器组织与血清,检测相关指标。结果与结论:①子宫组织大体观、苏木精-伊红染色和Masson染色结果:假手术组结构完整,内膜厚度适中,血管结构清晰。与假手术组相比,模型组子宫萎缩,结构不完整,内膜厚度、腺体数量明显减少,血管结构紊乱,纤维化增加;与模型组相比,经人脐血衍生物治疗后,子宫大小、结构、内膜厚度均有所恢复(均P<0.01),并且纤维化均减轻,其中以人脐血单个核细胞组恢复最为明显。在腺体数量增加方面,以人脐血富血小板血浆组效果最为显著(P<0.0001)。②子宫组织免疫组化与免疫荧光结果显示,与假手术组相比,模型组细胞增殖相关指标角蛋白9、波形蛋白,内膜容受性相关指标白血病抑制因子、整合素αγβ3,血小板内皮细胞黏附分子、碱性成纤维细胞生长因子、血管内皮生长因子表达均降低(均P<0.05)。与模型组相比,经人脐血衍生物治疗后以上指标均有显著性升高,人脐血衍生物各组比较:角蛋白9、血管内皮生长因子蛋白:人脐血单个核细胞组>人脐血间充质干细胞组>人脐血富血小板血浆组;波形蛋白、白血病抑制因子蛋白:人脐血间充质干细胞组>人脐血单个核细胞组>人脐血富血小板血浆组;整合素αγβ3蛋白:人脐血富血小板血浆组>人脐血间充质干细胞组>人脐血单个核细胞组;血小板内皮细胞黏附分子蛋白:人脐血富血小板血浆组>人脐血单个核细胞组>人脐血间充质干细胞组;碱性成纤维细胞生长因子蛋白:人脐血单个核细胞组>人脐血富血小板血浆组>人脐血间充质干细胞组。③Western blot检测显示,与假手术组相比,模型组白细胞介素6、白细胞介素1β和肿瘤坏死因子α蛋白表达水平显著升高(均P<0.001),经人脐血衍生物治疗后上述指标均降低(均P<0.05)。④ELISA检测显示,与假手术组相比,模型组抗缪勒氏管激素、雌二醇、孕酮水平降低,促卵泡生成素、促黄体生成素水平增加(除促黄体生成素外,均P<0.05)。经人脐血衍生物治疗后,性激素和抗缪勒氏管激素水平有一定程度恢复。⑤生育力实验显示,与假手术组相比,模型组受孕时间有所增加,产仔数明显降低(均P<0.05)。经人脐血衍生物治疗后,3组产仔数均有所增加(P<0.05),组间比较尚未发现明显差异。此研究初步表明人脐血单个核细胞对薄型子宫内膜有一定治疗效果,且人脐血富血小板血浆、人脐血单个核细胞和人脐血间充质干细胞分别在改善薄型子宫内膜再生功能、内膜容受性、血管生成、炎症调控及妊娠结局方面具有不同的优势与差异。
