J4 ›› 2010, Vol. 36 ›› Issue (2): 327-330.

• 基础研究 • 上一篇    下一篇

人脂肪源性干细胞成骨诱导过程中Ⅰ型胶原的表达

 王敏1, 孙新华2, 申玉芹3, 王景云1, 冯志远2   

  1. 1. 吉林大学口腔医院修复科| 吉林 长春 130021;2. 吉林大学口腔医院正畸科| 吉林 长春 130021;3. 吉林大学口腔医院牙周病科| 吉林 长春 130021
  • 收稿日期:2010-01-04 出版日期:2010-03-28 发布日期:2010-03-28
  • 通讯作者: 孙新华(Tel:0431-88796023,E-mail:xinhuasun8@163.com) E-mail:xinhuasun8@163.com
  • 作者简介:王 敏(1976-)|女|河南省洛阳市人|讲师|主治医师|在读医学博士|主要从事干细胞成骨诱导及其调控的研究。
  • 基金资助:

     吉林省科技厅白求恩医学专项基金资助课题 (200705344)

Expression of collagen type Ⅰ in human adipose-derived stem cells  during osteogenic differentiation and induction


 WANG Min1, SUN Xin-Hua2, SHEN Yu-Qin3, WANG Jing-Yun1, FENG Zhi-Yuan2   

  1. 1.Department of Prosthodontics,Stomatology Hospital,Jilin University,Changchun 130021;2. Department of Orthodontics,Stomatology Hospital,Jilin University,Changchun 130021;3. Department of Periodontics,Stomatology Hospital,Jilin University,Changchun 130021
  • Received:2010-01-04 Online:2010-03-28 Published:2010-03-28

摘要:

目的:研究人脂肪源性干细胞(hADSCs)成骨诱导后其合成和分泌Ⅰ型胶原能力的变化,探讨hADSCs作为组织工程骨的种子细胞的效能。方法:从人脂肪抽吸物中分离基质细胞,体外培养、扩增及鉴定。采用成脂诱导剂诱导hADSCs向脂肪细胞分化,采用油红O染色鉴定。采用成骨诱导剂诱导hADSCs向成骨细胞分化,将成骨诱导组分为0、7、14、21和28 d组,分别于0、7、14、21及28 d行茜素红染色检测矿化结节、Van Gieson胶原纤维特殊染色染胶原纤维、Ⅰ型胶原细胞免疫荧光染色检测细胞Ⅰ型胶原的表达。用FV Viewer 1.7软件对Ⅰ型胶原表达的强度进行定量检测。结果:hADSCs成脂诱导14 d后油红O染色阳性;hADSCs成骨诱导21 d茜素红染色阳性;成骨诱导21 d Van Gieson胶原纤维特殊染色阳性;hADSCs成骨诱导后Ⅰ型胶原染色21 d所有的细胞均阳性表达,28 d呈强阳性表达。统计结果显示:Ⅰ型胶原的表达从7 d开始,到28 d逐渐升高,7、14、21和28 d组与0 d组比较差异均有显著性(P<0.05)。结论:hADSCs成骨诱导后全部细胞呈现成骨表型,成骨能力强,可以作为骨种子细胞应用于临床骨再生治疗。

关键词: 脂肪源性干细胞;成骨诱导;多分化潜能;Ⅰ型胶原

Abstract:

Abstract:Objective To investigate the synthesis and secretion of collagen type Ⅰ(Col Ⅰ) in human adipose-derived stem cells(hADSCs) during osteogenic differentiation and induction,and approach the efficacy of hADSCs as seed cells for bone regeneration engineering.Methods hADSCs were separated,cultivated and amplified in vitro and were identified by cell phenotype detection. Multi-lineage differentiation potential was assessed by histological staining such as oil red O for lipid accumulation and alizarin red S for mineralization nodules during adipogenic and osteogenic induction. Osteogenic differentiation and induction group was divided into five subgroups:0,7,14,21 and 28 d groups. Alizarin red S staining,Van Gieson collagen fiber staining and Col Ⅰ immunofluorescence were used to detect the expression of Col Ⅰ. FV Viewer 1.7 software was used to analyze the data. Results The positive stainning of oil red O was found 14 d after adipogenic induction. For the osteogenic induction groups,the calcified nodules and the collagen fiber red staining were found in 21 d group. The expression of Col Ⅰ was found in 21 d group for all cells and  strongly positive expression in 28 d group. Compared with 0 d group,the expression of Col Ⅰ in osteogenic induction group was significantly increased from 7 d to 28 d(P<0.05). Conclusion During osteogenic induction of hADSCs,all cells have osteogenic capability and the hADSCs have outstanding capability of osteoinductive and osteogenesis. hADSCs may be useful in  clinical cell-based therapy for bone regeneration engineering.

Key words: adipose-derived stem cells;osteogenic  , induction;multi-lineage differentiation potential;collagen type Ⅰ

中图分类号: 

  • Q813.1