猪脂肪来源干细胞分离、培养及多向诱导分化研究  

Porcine adipose derived stem cells： isolation, characterization and multi-differentiation potential

作　　者：赵宝成[1] 赵博[1] 顾蓓[2] 徐慧民[3] 王振军[1] 

机构地区：[1]首都医科大学附属北京朝阳医院普外科,北京100020 [2]中国医学科学院基础医学研究所 [3]山东省潍坊市人民医院普外科

出　　处：《中华实验外科杂志》2011年第7期1099-1101,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察猪脂肪来源干细胞的分离、体外培养方法及基本生物学特性，探讨其多向诱导分化潜能。方法取猪颈部皮下脂肪组织，酶消化法获得脂肪干细胞，细胞免疫荧光及流式细胞技术检测细胞表面抗原，诱导其向脂肪细胞、软骨细胞和成骨细胞分化，油红染色、逆转录-聚合酶链反应（RT—PCR）和茜素红染色检测细胞诱导分化。结果成功培养出脂肪干细胞，流式细胞术检测CD44、CD105的阳性率分别为90．2％和99．2％，CD34、CD45的阳性率分别为0．3％和1．2％。成脂肪诱导3周后经油红O染色显示细胞内有脂滴形成，诱导成功率为93％，RT—PCR检测显示细胞能表达过氧化物酶体增殖物激活受体（PPAR-γ）；成软骨诱导4周后，RT—PCR检测结果显示细胞能表达软骨聚集蛋白聚糖（aggrecan）和Ⅱ型胶原；成骨诱导3周后，茜素红染色发现细胞出现钙结节，诱导成功率为94％。结论猪脂肪干细胞在体外具有向成脂肪、成软骨和成骨细胞分化的潜能，有望成为组织工程较理想的种子细胞之一。Objective To investigate the isolation, culture, biological characteristics and multidifferentiation of porcine adipose-derived stem cells （ADSCs） in vitro. Methods ADSCs were obtained from back subcutaneous adipose tissue of porcine by collagenase digestion and cultured in vitro. Flow cytometric analysis and eyto-immumofluorescence staining technique were carried out to detect the immunophenotypes of ADSCs. Reverse transcription-polymerase chain reaction （B.T-PCR） analysis and specific staining were used to evaluate the multipotential differentiation phenotype of ADSCs. Results The stem cells isolated from subcutaneous adipose tissue were positive for CD44 （90. 2% ） and CD105 （99. 2% ）, lacking in CD34 （0. 3% ） and CD45 （ 1.2% ）. After culture in mesenchymal stem cell adipogenic, chondrogenic and osteogenic differentiation basal media respectively for 3, 4 and 3 weeks, the cells were positive for oil red O staining （93%） and alizarin bordeaux staining （94%）, also the expression of peroxisome proliferator activated receptor （PPAR）-γ, collagen and aggrecan was detected in the differentiated cells. Conclusion The porcine ADSCs can differentiate towards adipocytes, chondrocytes and osteoblasts when culture in certain medium, and ADSCs can likely served as one of the most optimal autogenous ceil sources for tissue engineering.

关 键 词：猪 脂肪干细胞 免疫表型 诱导 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]