检索规则说明:AND代表“并且”;OR代表“或者”;NOT代表“不包含”;(注意必须大写,运算符两边需空一格)
检 索 范 例 :范例一: (K=图书馆学 OR K=情报学) AND A=范并思 范例二:J=计算机应用与软件 AND (U=C++ OR U=Basic) NOT M=Visual
作 者:胡佳辰 金岩 刘文佳 HU Jiachen;JIN Yan;LIU Wenjia(State Key Laboratory of Military Stomatology,National Clinical Research Center for Oral Diseases,Tissue Engineering Center,Third Affiliated Hospital,Air Force Medical University,Xi’an 710032,China;National&Local Joint Engineering Research Center of Biodiagnosis and Biotherapy,Precision Medicine Institute,The Second Affiliated Hospital,Xi’an Jiao Tong University,Xi’an 710004,China)
机构地区:[1]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,空军军医大学第三附属医院组织工程中心,陕西省西安市710032 [2]西安交通大学第二附属医院生物诊断与生物治疗国家地方联合工程研究中心,精准医疗研究院,陕西省西安市710004
出 处:《组织工程与重建外科》2021年第4期283-289,326,共8页Journal of Tissue Engineering and Reconstructive Surgery
基 金:国家重点研发计划(2016YFC1101400);国家自然科学基金面上项目(81970915)。
摘 要:目的探索肝脏脱细胞基质(dECM)微颗粒对3D培养HepG2细胞的肝相关功能状态的影响。方法通过肝脏门静脉灌流脱细胞液等成分,制得肝脏dECM。用组织学特殊染色、扫描电镜及免疫组织化学等方法鉴定脱细胞效果以及dECM成分。通过液氮冷冻研磨将dECM制备成微颗粒,通过差速离心分离不同粒径范围的微颗粒。利用超低吸附培养板进行HepG2细胞球状体3D培养,向培养基中加入dECM微颗粒,观察其对HepG2细胞球状体的形成及其肝相关功能状态的影响。利用ELISA、PCR检测相关蛋白的分泌和基因表达。结果制得的dECM无细胞结构,保留了细胞外基质的相关成分。在液氮研磨与差速离心分离后,可获得粒径在微米级的dECM微颗粒。dECM微颗粒可以促进HepG2细胞形成更致密的球状体;加入dECM微颗粒的HepG2球状体,细胞分泌白蛋白和尿素氮的水平增加(P<0.05);PCR结果表明,加入dECM微颗粒后,细胞的肝功能相关基因的表达水平显著提升(P<0.05)。结论肝脏脱细胞基质微颗粒可以促进HepG2细胞球状体的形成和HepG2细胞球状体的肝相关功能分化。Objective To explore the effects of liver decellularized extracellular matrix(dECM)microparticles on liver-related functional status of HepG2 cells cultured in 3D spheroids.Methods Liver dECM was prepared by perfusing detergent fluids through portal vein.The decellularization effect and the components of dECM were determined by histological special staining,scanning electron microscopy and immunohistochemistry.The dECM was prepared into microparticles by liquid nitrogen freeze grinding,and the microparticles with different particle size range were separated by differential centrifugation.3D HepG2 cell spheroids were cultured by ultra-low attachment culture plate and dECM microparticles were added into the culture medium to observe their effects on the formation of HepG2 cell spheroids and the functional status of HepG2 cell spheroids.The secretion and gene expression of related proteins were detected by ELISA and PCR.Results Obtained dECM retained the components of the extracellular matrix and no cellular components were observed.dECM particles with a particle size of several microns could be obtained after liquid nitrogen grinding and differential centrifugation.dECM particles could promote the formation of HepG2 cell spheroids.And the levels of albumin and urea nitrogen in culture supernatant were increased after the addition of dECM particles(P<0.05);PCR result also showed that the expression levels of genes related to liver function were increased after the addition of dECM particles(P<0.05).Conclusion Liver decellularized matrix microparticles can promote the formation of HepG2 cell spheroids and the differentiation of liver-related function of HepG2 spheroids.
关 键 词:细胞外基质 HEPG2细胞 细胞球状体 组织工程 3D培养
分 类 号:R318.1[医药卫生—生物医学工程]
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在链接到云南高校图书馆文献保障联盟下载...
云南高校图书馆联盟文献共享服务平台 版权所有©
您的IP:216.73.216.38