中国组织工程研究与临床康复2008，Vol.12Issue(14)：2780-2784,5.
骨髓间充质干细胞在膀胱脱细胞基质上的生长及分化
Culture and differentiation of bone marrow mesenchymal stem cells on bladder acellular matrix
摘要
Abstract
BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.关键词
膀胱脱细胞基质/骨髓间充质干细胞/生长分化/材料/组织工程膀胱分类
医药卫生引用本文复制引用
刘作强,黄海,黄健,林天歆,许可慰,郭正辉,江春,韩金利..骨髓间充质干细胞在膀胱脱细胞基质上的生长及分化[J].中国组织工程研究与临床康复,2008,12(14):2780-2784,5.基金项目
卫生部公益性行业科研专项(#03104)"腹腔镜技术在膀胱金切除及功能重建中的应用研究" （#03104）
广州市科技重点攻关项目资助(2005Z2-E0121)"腹腔镜膀胱切除-再造术治疗膀胱肿瘤的研究" （2005Z2-E0121）
卫生部部属医院临床重点项目(5010计划)(2007018)"膀胱癌腹腔镜根治术的应用和推广资助".Supported by:the Special Program for Scientific Research of Public Welfare of Ministry of Health,No.#03104 （5010计划）
the Tackle Key Programs in Science and Technology of Guangzhou City,No.2005Z2-E0121 （）
the Clinical Key Program of Hospital of Ministry of Health (5010 Program), No.2007018 （5010 Program）
