[1]黄宏,朱方强,孙宏振,等.脂肪来源干细胞体外成骨诱导和成脂诱导分化[J].第三军医大学学报,2008,30(13):1219-1222.
 HUANG Hong,ZHU Fang-qiang,SUN Hong-zhen,et al.Differentiation of swine adipose tissue-derived stromal cells into adipocytes and osteoblasts in vitro[J].J Third Mil Med Univ,2008,30(13):1219-1222.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第13期
页码:
1219-1222
栏目:
论著
出版日期:
2008-07-15

文章信息/Info

Title:
Differentiation of swine adipose tissue-derived stromal cells into adipocytes and osteoblasts in vitro
作者:
黄宏朱方强孙宏振冯帅南王正国朱佩芳张波
第三军医大学大坪医院野战外科研究所第四研究室,创伤、烧伤与复合伤国家重点实验室
Author(s):
HUANG Hong ZHU Fang-qiang SUN Hong-zhen FENG Shuai-nan WANG Zheng-guo ZHU Pei-fang ZHANG Bo
State Key Laboratory of Trauma, Burns and Combined Injury, Department 4, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, China
关键词:
干细胞脂肪组织脂肪来源干细胞脂肪细胞成骨细胞细胞分化
Keywords:
stem cells adipose tissue adipocytes osteoblasts differentiation
分类号:
R322.71; R329.21; R329.24
文献标志码:
A
摘要:
目的  探讨脂肪组织来源的多能干细胞(adipose tissue-derived stromal cells, ADSCs)的多向分化潜能。  方法  取小香猪腹部皮下脂肪组织,通过Ⅰ型胶原酶消化、离心等步骤分离培养ADSCs,经过原代培养和传代培养。分别加入成骨诱导剂和成脂诱导培养。经过倒置显微镜观察诱导后细胞形态变化,并通过Gomori 改良钙钴法、von  Kossa染色和油红O染色对其成骨细胞和脂肪细胞表型进行鉴定,进一步比较细胞的转化率。  结果   ADSCs呈成纤维细胞样贴壁生长,其经成骨、成脂诱导培养2周后形态、体积发生明显改变。Gomori 改良钙钴法染色显示其细胞质内富含碱性磷酸酶(alkaline phosphatase,ALP)颗粒,诱导后15 d和19 d,ALP阳性细胞分别为(25.0±7.5)%和(49.7±6.9)%,von Kossa染色表明聚集的细胞团中央能形成钙化结节;倒置显微镜,成脂诱导3、7、14 d后脂肪细胞转化率分别为(22.3±12.5)%、(49.6±7.1)%和(89.4±9.3)%。油红O染色可见细胞质内出现橙红色脂滴,14 d油红O染色阳性率高达(92.5±9.1)%。  结论  ADSCs经体外诱导培养后可向成骨细胞和脂肪细胞分化,并具有明显的成骨和成脂表型,表明ADSCs具有多向分化潜能。
Abstract:
Objective    To investigate the multilineage differentiation capacity of mesenchymal stem cells isolated and cultured from swine adipose tissue.     Methods    Adipose tissue-derived stromal cells (ADSCs) were obtained from adipose tissue of swine abdominal wall. The cells presented fibroblast-like in the culture medium. Adipose tissue was minced and digested with collagenase type Ⅰ. The obtained cells were plated and expanded in DMEM/F12 medium. Then the cells were cultivated in osteoblast-inducing culture medium, and osteoblast phenotype was assayed with Gomori and von Kossa staining. The passage cells were cultured in adipogenisis medium and stained with Oil Red O for identification. The cells were observed with inverted microscope each day.     Results    ADSCs grew as adherent cells and presented fibroblast-like in vitro, and could stably proliferate and be passed. Polygon-shaped cells were characterized by large size with a relatively large nucleus and plenty of cytoplasm after osteoblast-inducing. The cells changed from fibroblast-like into ellipse after adipogenisis-inducing cultivation. There were a lot of alkaline phosphatase (ALP) particles by Gomori staining. After induced for 15 and 19 d, positive cells for ALP were (25.0±7.5)%and (49.7±6.9)%, respectively. Von Kossa staining demonstrated the formation of mineralized nods in extracellular matrix. Under the inverted microscope, there existed significant lipid drops around the cell nucleus after adipogenisis-inducing cultivation for 2 d. The adipocytes rate on day 3, 7 and 14 was (22.3±12.5)%, (49.6±7.1)% and (89.4±9.3)%, respectively. After 14 d of adipocyte inducing, about (92.5±9.1)% cells displayed accumulation of lipid vacuoles, as detected by Red Oil O.     Conclusion    ADSCs isolated and cultured from swine adipose tissue can be induced to be osteo-inducing and adipogenisis, suggesting that the cells have the multilineage differentiation.

参考文献/References:

黄宏, 朱方强, 孙宏振, 等. 脂肪来源干细胞体外成骨诱导和成脂诱导分化[J]. 第三军医大学学报, 2008, 30(13):1219-1222.

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更新日期/Last Update: 2008-06-30