不同部位脂肪源性干细胞的生物学特性比较☆

摘要

背景：大鼠不同部位来源的脂肪源性干细胞在体外培养时的特性是否存在差异目前尚无定论。目的：比较同一只大鼠不同部位来源的脂肪源性干细胞在体外培养时的生长特性和成脂诱导分化能力的差异。方法：无菌操作下取 F344大鼠腹股沟及腹腔大网膜脂肪组织各5 mL，Ⅰ型胶原酶酶解法分离出脂肪源性干细胞，细胞计数后进行体外培养，观察其形态特征和生长状态，MTT 法测定不同部位细胞的倍增时间。取不同部位来源的第2代脂肪源性干细胞进行成脂诱导，诱导14 d 进行油红 O 染色，观察不同部位来源的脂肪源性干细胞的成脂分化能力。结果与结论：在同一只大鼠内脏大网膜脂肪获得的脂肪源性干细胞数目为(281±10)×107 L-1，明显多于腹股沟皮下脂肪的(85±5)×107 L-1(P <0.01)。从内脏大网膜脂肪与腹股沟皮下脂肪获得的脂肪源性干细胞分别于第5，6天进入指数增长期；第9，10天到达平台期；倍增时间为50 h 和60 h 左右。传代后的细胞生长分化活跃，呈成纤维细胞样，成脂诱导后，大网膜组织来源的脂肪源性干细胞的成脂诱导分化率明显高于腹股沟组织来源的脂肪源性干细胞[(38.90±2.86)%，(35.30±3.29)%，P <0.01]。可见同一只大鼠不同部位脂肪组织分离得到的脂肪源性干细胞数目不同，体外成脂诱导分化能力亦存在差异。%BACKGROUND: Whether the differences exist between adipose-derived stem cells isolated from different parts of rats when cultured in vitro has been poorly understood. OBJECTIVE: To compare the growth characteristics and adipogenic ability of adipose-derived stem cells isolated from different parts of rats. METHODS: Freshly isolated adipose-derived stem cells were obtained from 5 mL inguinal groove and greater omentum adipose tissue of F344 rats using type Ⅰ col agenase digestion method. Then, adipose-derived stem cells were counted and cultured in vitro. Morphological and growth characteristics of adipose-derived stem cells derived from the two sites were observed. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was utilized to examine the doubling time of adipose-derived stem cells from different parts. The passage 2 adipose-derived stem cells were induced adipogenical y. Fourteen days after being induced, the differentiated cells were stained with oil red O and the positive cells were counted. The adipogenic differentiation ability of adipose-derived stem cells from the different parts was assessed. RESULTS AND CONCLUSION: (1) The number of adipose-derived stem cells from the greater omentum fat tissue in the same group was (281±10)×107/L, which was significantly higher than that from the inguinal groove fat tissue [(85±10)×107/L] (P < 0.01). Adipose-derived stem cells from the greater omentum and inguinal groove fat tissue achieved the exponential growth period on days 5 and 6, respectively, and achieved the platform period on days 9 and 10, respectively. The corresponding doubling time was 50 hours and 60 hours, respectively. After being passaged, adipose-derived stem cells grew in fibroblast-like shape actively. The adipogenic differentiation rate of adipose-derived stem cells from the greater omentum fat tissue was higher than that from the inguinal groove fat tissue [(38.90±2.86)% vs. (35.30±3.29)%, P < 0.01]. This shows that the number and the adipogenic differentiation ability of adipose-derived stem cells from different parts of the same F344 rat are different.