人脂肪干细胞向淋巴管样内皮细胞分化的最佳条件  

Optimal induction conditions for adipose-derived stem cells differentiating into lymphatic endothelial cells

作　　者：陈小虎[1] 陈添和[1] 徐学战[1] 郭强[1] 

机构地区：[1]中山大学附属第一医院惠亚医院骨科,广东省惠州市516080

出　　处：《中国组织工程研究》2015年第32期5177-5181,共5页Chinese Journal of Tissue Engineering Research

基　　金：广州市医药卫生科技重点项目(201102A212004);惠州市科技局医疗卫生项目专题(20140802)~~

摘　　要：背景:作者所在课题组前期研究表明,脂肪干细胞在血管内皮生长因子C作用下,可以诱导分化为淋巴管样内皮细胞,经淋巴内皮细胞透明质酸受体1染色证实为淋巴管样内皮细胞,但其最佳诱导方案尚不明确。目的:探讨利用血管内皮细胞生长因子C(VEGF-C156s)诱导脂肪干细胞成为淋巴管样内皮细胞的最佳条件。方法:取健康成人脂肪组织,胰酶消化法获得脂肪组织来源的间质干细胞,通过流式细胞仪检测其表面标记,并进行成脂肪、成骨诱导等体外分化能力鉴定。取生长状态良好的第3代细胞,对照组加入低糖DMEM培养基,其余5组分别加入含25,50,100,200,300μg/L VEGF-C156s,分别观察诱导结果。结果与结论:成功分离纯化脂肪干细胞,在体外成功利用含VEGF-C156s、碱性成纤维生长因子等生长因子的诱导液将脂肪干细胞诱导为淋巴管内皮样细胞,对照组未见明显淋巴内皮细胞透明质酸受体1染色阳性细胞。诱导8 d后,25μg/L VEGF-C156s诱导后可见少数细胞染色阳性;50,100μg/L VEGF-C156s诱导后可见大量细胞淋巴内皮细胞透明质酸受体1阳性表达;200,300μg/L VEGF-C156s诱导会导致细胞大量死亡。说明以质量浓度为50μg/L的VEGF-C156s诱导8 d,是脂肪干细胞诱导分化为淋巴管样内皮细胞的最佳条件。BACKGROUND：Our previous studies have shown that adipose-derived stem cels under vascular endothelial growth factor C （VEGF-C） can be induced to differentiate into lymphatic endothelial cels that are confirmed by lymphatic vascular endothelial hyaluronan receptor-1 staining. However, its optimal induction program is not clear. OBJECTIVE：To investigate the best condition for the differentiation of adipose-derived stem cels into lymphatic endothelial cels under induction of VEGF-C156s. METHODS： Adipose tissues from healthy adults were colected to isolate adipose-derived mesenchymal stem cels using trypsin digestion method. Flow cytometry was employed to detect cel surface markers, andin vitro differentiation capacity was identified by adipogenic and osteogenic induction. Passage 3 cels at good growth state were selected and divided into six groups： cels in control group were cultured in low-glucose DMEM, and those in the rest five groups were treated with 25, 50, 100, 200, 300 μg/L VEGF-C156s, respectively. RESULTS AND CONCLUSION：Adipose-derived stem cels were successfuly obtained by trypsin digestion and purification, and then differentiated into lymphatic endothelial cels under the induction of VEGF-C156s, basic fibroblast growth factor and other growth factors. No cels were positive for lymphatic vascular endothelial hyaluronan receptor-1 in the control group. After 8 days of induction, few cels were positive in the 25 μg/L＆nbsp;VEGF-C156s group; a great amount of positive cels were visible in the 50 and 100 μg/L VEGF-C156s groups; 200 and 300 μg/L VEGF-C156s resulted in a large number of deaths in the cels. These findings indicate that it is optimal for adipose-derived stem cels to differentiate into lymphatic endothelial cels under 8-day induction of 50 μg/L VEGF-C156s.

关 键 词：干细胞 脂肪干细胞 淋巴管内皮样细胞 血管内皮细胞生长因子C 淋巴管内皮细胞透明质酸受体1 组织工程淋巴管 诱导 分化能力 

分 类 号：R394.2[医药卫生—医学遗传学]