兔膀胱移行上皮细胞与丝素蛋白膜相容性研究.pdfVIP

· 2l6· 南方医科大学学报(J South Med Univ) 2008；28(2) 兔膀胱移行上皮细胞与丝素蛋白膜相容性研究 刘春晓，廖云峰，李虎林，郑少波(南方医科大学珠江医院泌尿外科，广东 广州 510282) 摘要：目的探讨用于膀胱组织工程研究的移行上皮种子细胞的培养方法，检测丝素蛋白膜与膀胱移行上皮细胞的细胞 相容性，探讨构建泌尿系腔道组织工程器官的可能性。方法 (1)取兔膀胱移行上皮细胞体外培养，观察细胞形态变化及 生长、增殖过程，并进行免疫荧光鉴定；(2)分别设立空白组(无细胞)、阴性对照组(培养基)和实验组(丝素蛋白膜浸提 液) 取对数生长期的膀胱移行上皮细胞，lxlO4／ml接种于96孔板中，每组各5孔，于接种后24、72和 l20 h，通过MTT 法显色，并于酶标仪490nnl波长下检测吸光度值，计算相对增殖率，对丝素蛋白膜与膀胱移行上皮细胞的相容性进行 评估。结果 细胞在第9～10天融合形成单层，细胞形态呈多角形的铺路石子状，经免疫荧光鉴定证实为移行上皮细胞； 24、72和 l20 h实验组吸光度值分别为0．424___0．020、0．996___0．1l8和 1．285___0．048．阴性对照吸光度值分别为0．4l9± 0．030、1．105_+0．098和 1．228_+0．052，两组比较无明显差异(P0．05)。结论丝素蛋白膜与膀胱移行上皮细胞相容性良好， 具有作为泌尿系腔道组织工程支架的潜在运用价值。 关键词：组织工程；膀胱移行上皮细胞；丝素蛋白膜；细胞相容性 中图分类号：R329．29：Q8l3 文献标识码：A 文章编号：l673—4254(2008)02—02l6—03 Cytocompatibility of silk fibroin film with rabbit urinary bladder transitional epithelial cells ／n vitro LIU Chun-xiao，LIAO Yun-feng，LI Hu-lin，ZHENG Shao—bo Department of Urinary Surgery，Zhujiang Hospital，Southem Medical University，Guangzhou 5 1 0282，China Abstract：0bjective To obtain large quantities ofwell differentiated urinary bladder transitional epithelial cells for used as the seed cells in bladder tissue engineering． and evaluate the cytocompatibility of silk fibroin film with the transitional cells in vitro to assess the possibility of tissue．engineered urinary organ construction． Methods The urinary bladder transitional epithelial cells were isolated from the bladders of New Zealand rabbits and cultured in vitro as the seed cells． whose morphology was observed and the specific protein(cytokeratin)expression identified by immunofluorescence assay．nle cells were seeded in 96-well plates at l×l OVml and incubated with silk fibroin film leaching solution or culture medium(negative contro1)． MTT assay was performed to determine the eell proliferation rates of the wells