| 犬皮肤成纤维细胞的分离、培养及鉴定 |
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| 引用本文： | 辛毅,谭志敏,刘小慧. 犬皮肤成纤维细胞的分离、培养及鉴定[J]. 中国实验动物学报, 2010, 18(6): 480-483,I0008. DOI: 10.3969/j.issn.1005-4847.2010.06.007 |
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| 作者姓名： | 辛毅 谭志敏 刘小慧 |
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| 作者单位： | [1]首都医科大学附属北京安贞医院北京市心肺血管疾病研究所实验研究中心,北京100029 [2]首都医科大学附属北京安贞医院心内科,北京100029 |
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| 基金项目： | 北京市自然科学基金，国家自然科学基金 |
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| 摘 要： | 目的探索和建立适用于犬皮肤成纤维细胞的体外分离、培养及鉴定的技术方法。方法采用组织贴块培养法和胰蛋白酶、胶原酶Ⅰ联合消化法对犬皮肤成纤维细胞进行体外培养、传代。并对所培养的细胞进行倒置显微镜观察和苏木素-伊红染色,观察成纤维细胞形态,并对培养细胞行波形蛋白免疫荧光染色。结果倒置相差显微镜下可见长梭形细胞生长,苏木素-伊红染色可见细胞呈漩涡状、平行排列,第5代细胞免疫荧光检测波形蛋白(vimentin)表达阳性。结论建立了高效快速分离和稳定培养成纤维细胞的方法,为诱导犬心房纤维化提供了充足的种子细胞。
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| 关 键 词： | 成纤维细胞 细胞分离 细胞培养 鉴定 |
Isolation, Culture and Identification of Fibroblasts from Canine Skin |
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| XIN Yi,TAN Zhi-min,LIU Xiao-hui. Isolation, Culture and Identification of Fibroblasts from Canine Skin[J]. Acta Laboratorium Animalis Scientia Sinica, 2010, 18(6): 480-483,I0008. DOI: 10.3969/j.issn.1005-4847.2010.06.007 |
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| Authors: | XIN Yi TAN Zhi-min LIU Xiao-hui |
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| Affiliation: | 1.Department of Experimental Research Center of Beijing Institute of Heart Lung and Vascular Diseases,Beijing 100029,China;2.Department of Cardiology,Capital Medical University affiliated Beijing Anzhen Hospital,Beijing 100029,China) |
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| Abstract: | Objective To explore and set up the methodology of isolation,cultivation,and identification of skin fibroblasts in vitro.Methods Fibroblasts were released from dog skin by digestion of the dog skin with ype I collagenase and trypsin.The morphology and phenotype characterization of cultured fibroblasts was inspected by light microscopy after HE and immunocytochemical staining for vimentin.Results Fibroblasts exhibit spindle shape and arranged in swirl or parallel under the inverted phase-contrast microscope.Conclusions This study provides a technique of fast and efficient isolation and cultivation of skin fibroblasts,providing enough cell sources for dog atrial fibrosis research. |
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| Keywords: | Fibroblast Cell isolation Cell culture Cell identification |
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