人E 钙粘素融合蛋白基质对人骨髓间充质干细胞向肝细胞定向诱导分化的影响研究

doi:10.3969/j.issn.0258-8021. 2013. 06.010

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摘要人骨髓间充质干细胞（hBMSCs）源的肝细胞对于肝脏疾病的治疗和药物的开发具有巨大的应用潜力。利用基因工程技术构建了融合蛋白质粒，并通过真核细胞表达体系表达出了人E-钙粘素胞外域与免疫球蛋白Fc段的融合蛋白（hE-cadherin-Fc），并将其用于疏水材料的表面修饰仿生构建细胞-细胞间相互作用的细胞外微环境，检测其对hBMSCs向肝样细胞定向诱导分化的影响。诱导分化培养4周后，与组织培养板（TC-PS）、明胶（Gelatin）基质相比，hE-cadherin-Fc基质显著促进细胞表达ALB、CK18、HNF-4等肝细胞分化基因，并且细胞的糖原合成和吲哚青绿（ICG）摄取功能均显著提高。在hE-cadherin-Fc基质表面分化培养4周时，白蛋白和尿素合成的量为2.7 pg/细胞/d和36.7 pg/细胞/d，相对于TC-PS和Gelatin分别提高了42.1%和28.6%、57.5%和25.7%。综上所述，利用hE-cadherin-Fc基质化构建细胞外微环境，有利于促进人骨髓间充质干细胞向功能化肝细胞的定向分化。

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	徐建斌¹张妍¹李素华¹祝传顺¹
	2姚芳莲²赤池敏宏³杨军^1*

关键词 ：人骨髓间充质干细胞, 肝细胞, 分化, 融合蛋白, 人E-钙粘素

Abstract：Hepatocytes differentiating from human mesenchymal stem cells (hBMSCs) hold great potentials for cellbased approaches to liver diseases and drug development. In order to effectively expand and induce hepatic differentiation of hBMSCs, a fusion protein consisting of human E-cadherin extracellular domain and the immunoglobulin G Fc region (hE-cadherin-Fc) was biosynthesized and used as an extracellular matrix biomimicking epithelia cell adhesion junction. The effects of the hE-cadherin-Fc matrix on the hepatic differentiation of hBMSCs were studied in this paper. Compared with TC-PS and gelatin coated surfaces, the hE-cadherin-Fc matrix effectively stimulated the hBMSCs to progress toward the polygonal morphology of hepatocyte, and enhanced the differentiated cells expressing hepatocytespecific genes and livespecific functions. These results show hE-cadherin-Fc is a promising artificial extracellular matrix (ECM) for the hBMSCs differentiation via homophilic interaction of hE-cadherin and synergy between cell adhesion molecular and growth factors.

Key words： human mesenchymal stem cells hepatocytes differentiation fusion protein hE-cadherin

基金资助:国家自然科学基金（31070855）；国家重点基础研究发展计划（973计划）（2011CB606202）；天津市应用基础及前沿技术研究计划（10JCYBJC10400）

引用本文:

徐建斌¹张妍¹李素华¹祝传顺^1,2姚芳莲²赤池敏宏³杨军^{1*
. 人E 钙粘素融合蛋白基质对人骨髓间充质干细胞向肝细胞定向诱导分化的影响研究[J]. 中国生物医学工程学报, 2013, 32(6): 708-715.

XU Jian Bin¹ZHANG Yan¹LI Su Hua¹ZHU Chuan Shun^1,2YAO Fang Lian²TOSHIHIRO Akaike^{3 &n. Human ECadherinFc Fusion Protein Matrix Improving the Hepatic Differentiation of Human Mesenchymal Stem Cells. journal1, 2013, 32(6): 708-715.}}

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http://cjbme.csbme.org/CN/ 10.3969/j.issn.0258-8021. 2013. 06.010 或 http://cjbme.csbme.org/CN/Y2013/V32/I6/708

［1］Bianco P, Robey PG. Stem cells in tissue engineering ［J］. Nature, 2001, 414(1): 118-121.
［2］Kuo TK, Hung SP, Lee OK, et al. Stem cell therapy for liver disease: parameters governing the success of using bone marrow mesenchymal stem cells ［J］. Gastroenterology, 2008, 134(7): 2111-2121.
［3］吕静静，於学禅，沈秋霞，等. 干细胞在工程化组织构建与再生中的应用［J］. 中国生物医学工程学报，2013, 32(1): 99-113.
［4］Horwitz EM. Stem cell plasticity: the growing potential of cellular therapy ［J］. Arch Med Res, 2003, 34(6): 600-606.
［5］Chen Ye, Dong Xuejun, Zhang Guorong, et al. In vitro differentiation of mouse bone marrow stromal stem cells into hepatocytes induced by conditioned culture medium of hepatocytes ［J］. J Cell Biochem, 2007, 102(1): 52-63.
［6］王永红，王常勇，郭希民，等. 人骨髓间质干细胞体外扩增和向成骨细胞分化的实验研究［J］. 中国生物医学工程学报, 2002, 21(3): 251-274.
［7］Takenaga M, Fukumoto M, Hori Y. Regulated nodal signaling promotes differentiation of the definitive endoderm and mesoderm from ES cells ［J］. J Cell Sci, 2007, 120(12): 2078-2090.
［8］Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering ［J］. Nat Biotechnol, 2005, 23(1): 47-55.［9］Xu Jianbin, Zhu Chuanshun, Yang Jun, et al. hEcadherinFc fusion protein coated surface enhances the adhesionand proliferation of human mesenchymal stem cells ［J］. Colloids Surf B Biointerfaces, 2013, 109(1): 97-102.
［10］Discher DE, Mooney DJ, Zandstra PW. Growth factors, matrices, and forces combine and control stem cells ［J］. Science, 2009, 324(5935): 1673-1677.
［11］Guilak F, Cohen DM, Estes BT, et al. Control of stem cell fate by physical interactions with the extracellular matrix ［J］. Cell Stem Cell, 2009, 5(1): 17-26.
［12］Langer R, Tirrell DA. Designing materials for biology and medicine ［J］. Nature, 2004, 428(1): 487-492.
［13］Chen SS, Fitzgerald W, Zimmerberg J. Cellcell and cellextracellular matrix interactions regulate embryonic stem cell differentiation ［J］. Stem Cells, 2007, 25(3): 553-561.［14］丁亮华，罗光华，吴俊. 成人骨髓基质干细胞体外成骨细胞分化诱导［J］. 中国生物医学工程学报, 2006, 25(3): 333-337.
［15］Ito H, Steplewski A, Alabyeva T, et al. Testing the utility of rationally engineered recombinant collagenlike proteins for applications in tissue engineering ［J］. J Biomed Mater Res A, 2006, 76(3): 551-560.
［16］Hersel U, Dahmen C, Kessler H. RGD modified polymers: biomaterials for stimulated cell adhesion and beyond ［J］. Biomaterials, 2003, 24(24): 4385-4415.
［17］Nagaoka M, Jiang HL, Cho CS, et al. Application of recombinant fusion proteins for tissue engineering ［J］. Ann Biomed Eng, 2010, 38(3): 683-693.
［18］栾希英，段巧艳，张学光，等. 再生柞蚕丝素蛋白对人骨髓间充质干细胞体外扩增的支持作用［J］. 中国生物医学工程学报, 2007, 26(2): 276-281.
［19］Yu Meihua, Du Fengyi, Yang Jun, et al. Preparation and characterization of a VEGFFc fusion protein matrix for enhancing HUVEC growth ［J］. Biotechnol Lett, 2012, 34(9): 1765-1771.
［20］杨英，杨军. 含肝细胞生长因子(HGF)红色荧光融合蛋白基因质粒的构建［J］. 生物医学工程学杂志, 2009, 26(6): 1286-1290. 
［21］Stockinger A, Eger A, Wolf J, et al. Ecadherin regulates cell growth by modulating proliferationdependent b-catenin transcriptional activity ［J］. J Cell Biol, 2001, 154(6): 1185-1196.
［22］Georgopoulos NT, Kirkwood LA, Walker DC, et al. Differential regulation of growthpromoting signalling pathways by Ecadherin ［J］. PLoS One, 2010, 5(10): 13621-13634.
［23］Li L, Bennett SA, Wang LS. Role of Ecadherin and other cell adhesion molecules in survival and differentiation of human pluripotent stem cells ［J］. Cell Adh Migr, 2012, 6(1): 59-70.
［24］Yonemura S. Cadherinactin interactions at adherens junctions ［J］. Curr Opin Cell Biol, 2011, 23(5): 515-522.
［25］Yue Xiaoshan, Murakami Y, Tamai T, et al. A fusion protein NcadherinFc as an artificial extracellular matrix surface for maintenance of stem cell features ［J］. Biomaterials, 2010, 31(20): 5287-5296.
［26］Watt AJ, Garrison WD, Duncan SA. HNF4: A central regulator of hepatocyte differentiation and function ［J］. Hepatology, 2003, 37(6): 1249-1253.
［27］Clevers H. Wnt/βcatenin signaling in development and disease ［J］. Cell, 2006, 127(3): 469-480.