中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (20): 3675-3678.doi: 10.3969/j.issn.1673-8225.2011.20.016

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

人肾间质成纤维细胞体外增殖及胶原的合成

于国华1，孔令玲2，曲桂梅1，冯国营2，姚卫东1，林春华3   

  1. 青岛大学医学院附属烟台毓璜顶医院，1病理科，3器官移植中心，山东省烟台市  264001；2山东省滨州医学院烟台校区，山东省烟台市 264003
  • 收稿日期:2010-12-13 修回日期:2011-03-15 出版日期:2011-05-14 发布日期:2011-05-14
  • 通讯作者: 于国华★，男，1980年生，山东省潍坊市人，汉族，2006年泸州医学院毕业，硕士，主治医师，主要从事肾脏纤维化分子病理机制研究。 ygh0535@ hotmail.com
  • 作者简介:于国华★，男，1980年生，山东省潍坊市人，汉族，2006年泸州医学院毕业，硕士，主治医师，主要从事肾脏纤维化分子病理机制研究。 ygh0535@ hotmail.com
  • 基金资助:

    课题受山东省中医药科技发展计划项目(2009222)资助, 课题名称：褐藻多糖硫酸酯对人肾小管上皮细胞间质转分化影响的实验研究。

In vitro proliferation and collagen synthesis of human renal interstitial fibroblasts

Yu Guo-hua1, Kong Ling-ling2, Qu Gui-mei1, Feng Guo-ying2, Yao Wei-dong1, Lin Chun-hua3   

  1. 1Department of Pathology, 3Department of Transplant Center, Affiliated Yantai Yuhuangding Hospital, Medical College of Qingdao University, Yantai  264001, Shandong Province, China；2Yantai Campus, Binzhou Medical University, Yantai  264003, Shandong Province, China
  • Received:2010-12-13 Revised:2011-03-15 Online:2011-05-14 Published:2011-05-14
  • Contact: Yu Guo-hua★, Master, Attending physician, Department of Pathology, Affiliated Yantai Yuhuangding Hospital, Medical College of Qingdao University, Yantai 264001, Shandong Province, China ygh0535@hotmail.com
  • About author:Yu Guo-hua★, Master, Attending physician, Department of Pathology, Affiliated Yantai Yuhuangding Hospital, Medical College of Qingdao University, Yantai 264001, Shandong Province, China ygh0535@hotmail.com
  • Supported by:

    the Science and Technology Development Plan for Traditional Chinese Medicine in Shandong Province, No. 2009222*

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摘要：

背景：肾间质成纤维细胞体外研究时间点的选取多在72 h以内，但对于体外长时间动态观察培养过程中成纤维细胞增殖及胶原分泌规律少见报道。
目的：观察人肾间质纤维化来源的成纤维细胞体外增殖及胶原蛋白合成的规律。
方法：采用组织块培养法获取、纯化并鉴定肾间质纤维化来源的人肾间质成纤维细胞，选取第3 代对数生长期细胞用于实验。MTT 比色法及Masson三色染色结合图像分析法分别检测体外培养第1，3，5，7，9 天细胞的吸光度值和成纤维细胞胶原蛋白染色阳性细胞的平均吸光度值。
结果与结论：肾间质成纤维细胞体外生长曲线呈“S”形，体外培养第3~5天为细胞倍增时间，随培养时间延长，胶原蛋白由线状分布转变成片状分布，其中以第3~5天增长最快(P < 0.05)。结果证实，肾间质成纤维细胞在体外培养第3~5天增殖能力和胶原合成活性最强。

关键词: 肾间质纤维化, 成纤维细胞, 增殖, 胶原, 体外培养, 组织构建

Abstract:

BACKGROUND: The time points for in vitro studies about renal interstitial fibroblasts are mainly selected within 72 hours. However, there are rare reported about proliferation and collagen synthesis of renal interstitial fibroblasts cultured in vitro for a long term.
OBJECTIVE: To investigate the pattern of proliferation and collagen synthesis of human renal interstitial fibroblasts in vitro.
METHODS: human renal interstitial fibroblasts were obtained, purified and identified. The third passage cells were used in experiment. Cell proliferation was determined by MTT method and collagen synthesis was observed using Masson trichrome staining accompanied with image analysis at the 1st, 3rd, 5th, 7th and 9th days after culture in vitro.
RESULTS AND CONCLUSION: The growth curve of renal interstitial fibroblasts was shown as “S” and doubling time was from the 3rd to 5th day in vitro; with the culturing time extending, the distribution of collagen protein changed from line into lamellar and the quantity of collagen protein increased obviously from the 3rd to 5th day (P < 0.05). It is indicated that the biological activity of renal interstitial fibroblasts is the most productive from the 3rd to 5th day, which might be an important target during the course of pharmacological experiment in vitro.

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