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| 1 | 小鼠骨髓间充质干细胞的分离、培养新方法显示文摘目的探讨一种新的小鼠骨髓间充质干细胞的分离、培养方法。方法分离小鼠3 d乳鼠双后肢,剔除肌肉筋膜组织,剪碎后接种于含10%胎牛血清的α-MEM中,观察细胞形态学变化,采用CCK8检测细胞增殖能力,绘制细胞生长曲线,采用流式细胞技术鉴定细胞表面抗原,并进行多向分化潜能鉴定。结果原代分离培养的小鼠骨髓间充质干细胞呈长梭形,从骨片周围爬出,传代后,细胞形态一致,生长良好;绘制的细胞生长曲线呈'S'型,流式细胞表型鉴定结果显示,培养的细胞高表达CD90、CD29,低表达CD11b、CD45,成脂油红O染色和成骨茜素红染色均呈阳性。结论采用小鼠3 d乳鼠骨片法能够成功培养出骨髓间充质干细胞,骨髓间充质干细胞可作为肝脏组织工程的种子细胞。 | 徐丽娟 张云巍 王淑芳 阎丽 | 2017 | 肝脏2017，22，3： | 7 |
| 2 | Wnt/β-catenin信号调控EMT水平对小鼠胚胎干细胞向肝脏组织结构分化的影响显示文摘目的在前期体外诱导小鼠胚胎干细胞(ESC)向肝系细胞分化基础上,从细胞上皮间质转化(EMT)角度研究ESC分化为肝脏组织结构的可行性。方法选用Balb/c小鼠ESC,首先利用肝细胞生长因子(HGF)、成纤维细胞生长因子(FGF)等体外诱导ESC向肝系细胞方向分化,然后在分化中期(13 d)、成熟期(17 d)两个时间点动态抑制细胞Wnt/β-catenin信号以降低其EMT水平。最后在分化系统中观察分化细胞保持三维组织结构生长情况,检测其中肝系细胞和血管标志物白蛋白(ALB)、血管内皮生长因子(VEGFR)等表达情况,综合判断有无肝脏组织形成,并研究该过程中EMT水平变化的规律和机制。结果ESC分化过程中,分化前期实验组EMT水平与对照组差别不明显,分化中、后期实验组EMT水平较对照组明显降低,分化第18天和20天,实验组上皮钙黏蛋白(E-cad)基因mRNA相对表达水平分别为0.61±0.15和0.47±0.05,对照组分别为0.07±0.05和0;实验组的ALB/AFP/CK8/CK19表达水平在同期内较对照组普遍偏高,标志物CD31/VEGFR1在分化中后期较对照组下降缓慢。在ESC培养上清液中,实验组ALB于第11天可检测到,浓度为(0.32±0.02)mg/L,而对照组ALB为(0.19±0.05)mg/L;实验组尿素于第13天开始检测到,浓度为(8.7±1.0)μmol/L,对照组尿素浓度为(3.1±1.2)μmol/L。ALB和尿素合成在ESC分化系统培养上清中浓度随分化时间延长而明显升高,并且实验组的ALB、尿素浓度值在相同的时间段内明显多于对照组。另外,实验组分化细胞可保持三维组织状态生长,对照组分化细胞最终呈单细胞状态。实验组可检测到肝系细胞标志物以及血管细胞标志物的表达,免疫荧光结果显示类肝细胞和血管结构紧密层状排列,HE染色提示有类肝小叶结构形成,而对照组无血管成分标志物表达。结论ESC向肝系细胞分化中,在分化中、后期降低EMT水平,可有效促进ESC向肝脏组织结构分化。 | 廖文尉 章诚 刘芙蓉 凌象超 蔡炳 朱晓峰 何晓顺 胡安斌 | 2018 | 中华医学杂志2018，98，30： | 7 |
| 3 | Adipose-derived stromal cells resemble bone marrow stromal cells in hepatocyte differentiation potential in vitro and in vivo显示文摘AIM To investigate whether mesenchymal stem cells(MSCs) from adipose-derived stromal cells(ADSCs) and bone marrow stromal cells(BMSCs) have similar hepatic differentiation potential.METHODS Mouse ADSCs and BMSCs were isolated and cultured. Their morphological and phenotypic characteristics, as well as their multiple differentiation capacity were compared. A new culture system was established to induce ADSCs and BMSCs into functional hepatocytes. Reverse transcription polymerase chain reaction, Western blot, and immunofluorescence analyses were performed to identify the induced hepatocytelike cells. CM-Dil-labeled ADSCs and BMSCs were then transplanted into a mouse model of CCl4-induced acute liver failure. fluorescence microscopy was used to track the transplanted MSCs. Liver function was tested by an automatic biochemistry analyzer, and liver tissue histology was observed by hematoxylin and eosin(HE) staining.RESULTS ADSCs and BMSCs shared a similar morphology and multiple differentiation capacity, as well as a similar phenotype(with expression of CD29 and CD90 and no expression of CD11 b or CD45). Morphologically, ADSCs and BMSCs became round and epithelioid following hepatic induction. These two cell types differentiated into hepatocyte-like cells with similar expression of albumin, cytokeratin 18, cytokeratin 19, alpha fetoprotein, and cytochrome P450. fluorescence microscopy revealed that both ADSCs and BMSCs were observed in the mouse liver at different time points. Compared to the control group, both the function of the injured livers and HE staining showed significant improvement in the ADSC-and BMSC-transplanted mice. There was no significant difference between the two MSC groups.CONCLUSION ADSCs share a similar hepatic differentiation capacity and therapeutic effect with BMSCs in an acute liver failure model. ADSCs may represent an ideal seed cell type for cell transplantation or a bio-artificial liver support system. | Li-juan Xu Shu-fang Wang De-Qing Wang Lian-jun Ma Zheng Chen Qian-Qian Chen Jun Wang Li Yan | 2017 | World Journal of Gastroenterology2017，23，38： | 7 |
| 4 | 非酒精性脂肪性肝病体外模型的研究进展显示文摘非酒精性脂肪性肝病发病率逐年上升,其治疗手段有限,对其发病机制的研究是当前热点。为了更好地阐明其发病机制,迫切需要开发先进、安全、有效的体外模型或体内模型,以了解并制订针对该疾病的治疗策略。本文回顾了目前常用于非酒精性脂肪性肝病临床前研究的体外模型,并讨论了其各自的优缺点,为研究非酒精性脂肪性肝病发病机制及治疗提供理论依据。 | 卢晓臣 韩红梅 | 2022 | 临床肝胆病杂志2022，38，1： | 2 |
| 5 | Derivation and applications of human hepatocyte-like cells显示文摘Human hepatocyte-like cells (HLCs) derived from human pluripotent stem cells (hPSCs) promise a valuable source of cells with human genetic background, physiologically relevant liver functions, and unlimited supply. With over 10 years’ efforts in this field, great achievements have been made. HLCs have been successfully derived and applied in disease modeling, toxicity testing and drug discovery. Large cohorts of induced pluripotent stem cells-derived HLCs have been recently applied in studying population genetics and functional outputs of common genetic variants in vitro. This has offered a new paradigm for genomewide association studies and possibly in vitro pharmacogenomics in the nearly future. However, HLCs have not yet been successfully applied in bioartificial liver devices and have only displayed limited success in cell transplantation. HLCs still have an immature hepatocyte phenotype and exist as a population with great heterogeneity, and HLCs derived from different hPSC lines display variable differentiation efficiency. Therefore, continuous improvement to the quality of HLCs, deeper investigation of relevant biological processes, and proper adaptation of recent advances in cell culture platforms, genome editing technology, and bioengineering systems are required before HLCs can fulfill the needs in basic and translational research. In this review, we summarize the discoveries, achievements, and challenges in the derivation and applications of HLCs. | Shuang Li Shi-Qian Huang Yong-Xu Zhao Yu-Jie Ding Dan-Jun Ma Qiu-Rong Ding | 2019 | World Journal of Stem Cells2019，11，8： | 1 |
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