脂肪干细胞体外培养特性及成脂成软骨分化研究

Study on the properties,adipogenic and chondrogenic differentiation of adipose-derived stem cells in vitro

目的:分析脂肪干细胞体外培养的特性和体外向成脂细胞和成软骨细胞分化的情况。方法:采集成年兔腹部脂肪组织,体外分离培养脂肪干细胞,并进行成脂细胞和成软骨细胞的体外诱导分化,采用油红O染色证明成脂细胞分化,Ⅱ型胶原免疫组织化学染色和Alcian Blue染色证明成软骨细胞分化。结果:脂肪干细胞形态为菱形,细胞可稳定传代,各代次间无形态上的差异。经油红O染色发现,脂肪干细胞可形成阳性脂滴,表明具有成脂分化能力。经Ⅱ型胶原免疫组织化学染色和Alcian Blue染色后发现,脂肪干细胞可形成Ⅱ型胶原和硫酸蛋白聚糖,表明具有成软骨分化能力。结论:脂肪干细胞具有成脂成软骨分化能力,可成为脂肪和软骨组织工程研究的种子细胞。

Objective: To analyze the properties,adipogenic and chondrogenic differentiation of adipose-derived stem cells in vitro.Methods: The abdominal adipose tissue of rabbit was harvested. The adipose-derived stem cells were isolated and cultured,and induced to adipogenic cells and cartilage cells in vitro. The adipogenic and chondrogenic differentiation of adipose-derived stem cells were identified using the oil red O staining,and type Ⅱ collagen immunohistochemical staining and Alcian Blue staining,respectively.Results: The adipose-derived stem cell was prismatic cells,and could stably passage. There was not the morphology difference in each generation cells. The results of oil red O staining found that the adipose-derived stem cells could form the positive lipid droplets,which demonstrated that the cells had adipogenic differentiation ability. The type Ⅱ collagen immunohistochemical staining and Alcian Blue staining showed that the adipose-derived stem cells could secrete the type Ⅱ collagen and sulfated proteoglycans,which demonstrated that the cells had the chondrogenic differentiation ability. Conclusions: The adipose-derived stem cells have the adipogenic and chondrogenic differentiation abilites which can become the seed cell of fat and cartilage tissue engineering.