[1]赵佐芳,王宁,王大庆,等.眼睑基底细胞癌相关成纤维细胞与正常成纤维细胞形态与生长增殖特性差异[J].眼科新进展,2016,36(4):327-330.[doi:10.13389/j.cnki.rao.2016.0088]
　ZHAO Zuo-Fang,WANG Ning,WANG Da-Qing,et al.Differences in cellular morphology and proliferation of eyelid basal cell carcinoma associated fibroblasts and normal fibroblasts[J].Recent Advances in Ophthalmology,2016,36(4):327-330.[doi:10.13389/j.cnki.rao.2016.0088]
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眼睑基底细胞癌相关成纤维细胞与正常成纤维细胞形态与生长增殖特性差异
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年4期
页码:
327-330
栏目:
实验研究
出版日期:
2016-04-05

文章信息/Info

Title:
Differences in cellular morphology and proliferation of eyelid basal cell carcinoma associated fibroblasts and normal fibroblasts
作者:
赵佐芳; 王宁; 王大庆; 兰长骏
637000　四川省南充市，川北医学院附属医院眼科，川北医学院眼视光学系
Author(s):
ZHAO Zuo-Fang ; WANG Ning ; WANG Da-Qing ;  LAN Chang-Jun
Department of Ophthalmology , Affiliated Hospital of North Sichuan Medical College, Department of Eye Optical of North Sichuan Medical College , Nanchong 637000 , Sichuan Province , China
关键词:
眼睑基底细胞癌; 成纤维细胞; 原代培养; 生物学特性
Keywords:
eyelid basal cell carcinoma;  fibroblasts ;  primary culture ;  biological characteristics
DOI:
10.13389/j.cnki.rao.2016.0088
文献标志码:
A
摘要:
目的　对眼睑基底细胞癌相关成纤维细胞（ｂａｓａｌｃｅｌｌｃａｒｃｉｎｏｍａ-ａｓｓｏｃｉａｔｅｄｆｉｂｒｏｂｌａｓｔｓ，ＢＣＡＦｓ）及眼睑正常成纤维细胞（ｎｏｒｍａｌｆｉｂｒｏｂｌａｓｔｓ，ＮＦｓ）进行分离、培养、纯化和鉴定，初步研究ＢＣＡＦｓ与ＮＦｓ细胞形态、生长增殖等生物学性状的差异。方法　用组织块贴壁法培养获得原代眼睑ＢＣＡＦｓ和眼睑ＮＦｓ，通过胰蛋白酶差速消化法和反复传代法进行细胞的分离、纯化；通过倒置相差显微镜对照观察ＢＣＡＦｓ和ＮＦｓ细胞形态，应用免疫细胞化学染色检测细胞表达的特异性蛋白鉴定ＢＣＡＦｓ，ＭＴＴ法检测细胞生长增殖特性。结果　在接种后３ｄ便可见原代ＢＣＡＦｓ从组织块爬出，呈放射状生长，约７ｄ铺满瓶底。ＮＦｓ生长缓慢，接种后４ｄ才见稀疏的ＮＦｓ在组织块周围生长，约１１ｄ铺满瓶底。传代后的ＢＣＡＦｓ在形态上未发生明显改变，但生长速度增快，与ＮＦｓ在形态结构特征和生长方式上存在差异；眼睑ＢＣＡＦｓ和ＮＦｓ的免疫细胞化学染色，前者波形蛋白、α-平滑肌肌动蛋白、成纤维细胞活化蛋白呈阳性，细胞角蛋白呈阴性，后者仅波形蛋白呈阳性，角蛋白、α-平滑肌肌动蛋白、成纤维细胞活化蛋白均呈阴性；与ＮＦｓ相比，眼睑ＢＣＡＦｓ生长增殖较快，倍增时间较短。结论　与眼睑ＮＦｓ相比，ＢＣＡＦｓ在形态结构、蛋白表达、生长方式等方面均有差异，且增殖能力强。
Abstract:
Objective To separate , culture , purify and identify eyelid basal cell carcinoma associated fibroblasts ( BCAFs) and eyelid normal fibroblasts ( NFs) , and research their biological character differences in cellular morphology and proliferation. Methods Primary eyelid BCAFs and NFs were cultivated by tissue block adhering wall method and purified by subculturing cells with pancreatic enzyme differential digestion. The morphology of BCAFs and NFs was observed by inverted phase contrast microscope. BCAFs was identified by immunocytochemical staining to detect cell specific protein expression. Cell growth and proliferation was evaluated by MTT. Results Primary BCAFs grew out from the tissue section at 3 days after cultured, BCAFs were long spindle , and spread all the bottom of bottle at 7 days ; NFs grew slowly , out from the tissue section at 4 days after cultureing , and spread all the bottom of bottle at 11 days. The morphous of passaged BCAFs had no obvious change , grow fast with disordered arrangement and overlap growth phenomenon, and had the differences in cellular morphology and proliferation with NFs. BCAFs had hiff expression of vimentin ,dÒ»smooth muscle actin and fibroblast activation protein but no expression of cytokeratin, however NFs only had high expression of vimentin and no expression of cytokeratin .d¡ªsmooth muscle actin and fibroblast activation protein. The growth and proliferation of BCAFs were fast than those of NFs, and the doubling time of BCAFs was shorter. Conclrision Compared with eyelid NFs , the eyelid BCAFs have many differences in morphology , expression of protein and growth way , the proliferation ability of BCAFs is stronger.

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更新日期/Last Update: 2016-04-01