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| 组织块结合酶消化法培养婴幼儿血管瘤内皮细胞 |
| 廖洪跃1,邢新1*,欧阳天祥2,郭伶俐1,李军辉1,薛春雨1,袁斯明3,李鸣1 |
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| (1.第二军医大学长海医院整形外科,上海 200433*2.上海交通大学医学院新华医院整形外科,上海 200092*3.南京军区总医院烧伤整形科,南京 210002) |
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| 摘要: |
| 目的:探讨血管瘤内皮细胞体外培养的方法并观察其生物学特性。方法:收集新鲜手术切除的婴幼儿血管瘤标本,采用组织块结合酶消化法培养内皮细胞,免疫组化EnVision法对培养的细胞进行鉴定;流式细胞仪FITC-CD34检测内皮细胞纯度;相差显微镜下观察血管瘤内皮细胞的生物学特性。结果:共收集血管瘤标本14例,有8例成功培养出内皮细胞,内皮细胞呈现两种形态:多角形和梭形,经免疫组化鉴定为内皮细胞;培养的细胞中CD34＋细胞数为76.28%;血管瘤内皮细胞有明显的“管腔形成”。结论:组织块结合酶消化法能成功培养较纯的婴幼儿血管瘤内皮细胞,培养的内皮细胞具有某些生物学特性。 |
| 关键词: 血管瘤 婴幼儿 内皮细胞 细胞培养 组织块结合酶消化法 |
| DOI：10.3724/SP.J.1008.2009.0147 |
| 投稿时间：2008-04-30修订日期：2008-12-03 |
| 基金项目:国家自然科学基金(30571928). |
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| Explant combined with trypsin-digestion for culture of endothelial cells from infantile hemangiomas |
| LIAO Hong-yue 1, XING Xin 1*, OUYANG Tian-xiang 2, GUO Ling-li 1, LI Jun-hui 1, XUE Chun-yu 1, YUAN Si-ming3, LI Ming1 |
| (1.Department of Plastic Surgery,Changhai Hospital,Second Military Medical University,Shanghai 200433,China*2.Department of Plastic Surgery,Xinhua Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200092*3.Department of Burn and Plastic Surgery,Nanjing General Hospital,PLA Nanjing Military Area Command,Nanjing 210002) |
| Abstract: |
| Objective:To explore a novel method for cultivation of the endothelial cells(EC) from infantile hemangiomas(IH) in vitro and observe the biological character of cultured endothelial cells.Methods:Fresh operative specimens were obtained from infantile hemangiomas.The endothelial cells of IH were cultured by explant combined with trypsin-digestion technique.Immunohistochemical staining of EnVision method was carried out to identify the cultured cells.The purity of endothelial cell was examined by flow cytometry analysis of FITC-CD34.The biologic characters of endothelial cells were observed under inverted phase contrast microscope.Results:Endothelial cells were successfully cultured from 8 IH specimens of all the 14 explants.The morphology of cultured endothelial cells included two types:polygonal cells and fusifourm cells.The cultured cells were homogenously positive for EC markers-vWF or CD34,indicating their EC origin.Tube formation was found in endothelial cells of IH.The proportion of CD34+ cells was 76.28% in cultured endothelial cells as detected by flow cytometry analysis.Conclusion:ECs can be successfully isolated and cultured from infantile hemangiomas by explant combined with trypsin-digestion technique; the cultured cells have some characters of endothelial cells. |
| Key words: hemangioma infantile endothelial cells cell culture explant combined with trypsin-digested technique |
