SD大鼠脂肪干细胞的体外分离培养及实验研究

Isolation and Culture in Vitro Stromal Cells from SD Rat Adipose Tissue and Its Experimental Study

目的探索从脂肪组织中分离、培养SD大鼠脂肪干细胞(Adipose tissue-derivedstromal cells,ADSCs)的方法,同时观察其生物学特性。方法取SD大鼠腹股沟区皮下脂肪组织,0.1%Ⅰ-型胶原酶消化分离、培养ADSCs,流式细胞术测定CD44、CD45和CD49d抗原的表达,MTT比色法测定细胞生长活力,诱导培养基向脂肪细胞定向分化,Oil Red O染色鉴定。结果分离出的细胞CD44表达阳性,CD49d表达弱阳性和CD45表达阴性,其生长曲线呈倒"S"形,成脂诱导培养基定向诱导分化,经Oil Red O染色呈红色。结论本次实验所分离出来的SD大鼠ADSCs在体外具有生长稳定,增殖较快并能诱导分化的特点。

Objective To explore the method of isolating and culturing adipose tissue-derived stromal cells from SD rat adipose tissue, and observe its biological characteristics in vitro. Methods The subcutaneous adipose tissue was obtained from iliae region of rat under the aseptic condition and was digested with 0. 1% I-eollagenase. The mononuelear cells were collected and cultured in DMEM with 10% fetal bovine serum. The expression of CD44 and CD49d were assayed by using flow eytometry. The cell growth vigor was tested by MTT ehromatometry, the passed cells were exposed to a defined medium for adipoeyte differentiation, and the propertiesof adipoeyte was identified with oil red O staining. Results The fibro-liked cells isolated from the rat＇s adipose tissue could rapidly expand. The results showed that CD44 showed positive, CD49d weakly positive and CD45 negative on the cell membrane by flow eytometer. The growth curve was opposite-like ＂S＂shape. The passed cells were exposed to a defined medium for adipoeyte differentiation, and then can be stained with oil red. Condes＇ion The ADSCS of SD rats isolated in this trial are characterized by stable growth, quick proliferation and induced differentiation.