两种培养基诱导人和大鼠脂肪干细胞向脂肪细胞分化的比较

Comparison of adipogenic differentiation ability of adipose stem cells derived from humans and rats using two kinds of adipogenic culture media

目的比较两种培养基诱导人和大鼠脂肪干细胞(adipose-derived stem cell,ADSC)的成脂分化能力,探寻更为适宜的ADSC成脂诱导方案。方法提取3例临床患者的脂肪干细胞(h ADSCs)及出生6周大鼠的脂肪干细胞(r ADSCs),将h ADSCs、r ADSCs分别接种于六孔板,分为未诱导组、成脂诱导I组和II组,分别使用普通培养基、成脂诱导培养基I和II,培养10 d后以油红O染色,镜下观察及检测490 nm吸光度值(A490),比较脂滴形成情况。结果未诱导的h ADSCs和r ADSCs均呈成纤维细胞样生长,成脂诱导培养3 d后出现脂滴;油红O染色显示,成脂诱导组脂滴呈橘红色,h ADSCs和r ADSCs的成脂诱导II组形成的脂滴均明显大于诱导I组,统计学分析显示h ADSCs的成脂诱导II组测得的A490明显高于诱导I组(P<0.01);而两组的r ADSCs的A490无明显差异(P>0.05)。结论 h ADSCs和r ADSCs在两种诱导培养基中均可成脂分化,但成脂诱导II组优于成脂诱导I组。

Objective To find a more appropriate method of adipogenic induction for adipose stem cells derived from humans and rats by comparing their adipogenic differentiation ability using two kinds of adipogenic culture media.Methods The human adipose stem cells（ h ADSCs） were extracted from lipoaspirates of 3 donors in the clinic,and the rat adipose stem cells（ r ADSCs） were obtained from adipose tissues of 6-week old rats. The cells were plated into two 6-well plates,and were divided into 3 groups： the negative control group,adipogenic induction group I and group II,respectively,two wells in each group. The control medium,adipogenic induction medium I and medium II were added into the cells of different groups,respectively. After induction for 10 days,the adipogenic differentiation ability of cells was assessed under microscope with oil red O staining and detecting the optical density value of 490 nm（ A 490 nm） to compare the lipid droplet formation in the cells. Results The h ADSCs and r ADSCs showed a fibroblast-like growth. Positive oilred O staining cells showed orange lipid in the cells of adipogenic induction group from the third day of culture. The amount of lipid in cells induced by adipogenic induction medium II was higher than that in cells induced by adipogenic induction medium I,the A490 nm optical density of hA DSCs induced by adipogenic induction medium II was significantly higher than that induced by adipogenic induction medium I（ P〈0. 01）,but there was no significant difference between the rA DSCs induced by adipogenic induction medium I and II（ P〉0. 05）. Conclusions Both hA DSCs and rA DSCs can be induced to adipogenic differentiation using the two kinds of induction media,however,the induction of adipogenic differentiation ability of the adipogenic induction medium II is stronger than that of the adipogenic induction medium I.