人脂肪源性干细胞成骨诱导过程中Ⅰ型胶原的表达

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

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. 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.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/Abstract

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

王敏, 孙新华, 申玉芹, 王景云, 冯志远. 人脂肪源性干细胞成骨诱导过程中Ⅰ型胶原的表达[J]. J4, 2010, 36(2): 327-330.

WANG Min, SUN Xin-Hua, SHEN Yu-Qin, WANG Jing-Yun, FENG Zhi-Yuan. Expression of collagen type Ⅰ in human adipose-derived stem cells 　during osteogenic differentiation and induction
[J]. J4, 2010, 36(2): 327-330.