脂肪组织来源干细胞与脂肪瘤间充质干细胞特性的比较研究  

The differences between adipose tissue derived stem cells and lipoma mesenchymal stem cells in characteristics

作　　者：钱奕炜[1] 高建华[1] 鲁峰[1] 郑旭东[1] 

机构地区：[1]南方医科大学南方医院整形外科,广州510515

出　　处：《中华整形外科杂志》2010年第2期125-132,共8页Chinese Journal of Plastic Surgery

摘　　要：目的 比较体外培养的脂肪来源干细胞（adipose-derived stem cells,ASCs）与脂肪瘤间充质干细胞（lipoma-derived mesenchymal stem cells,LMSCs）的生物特性,以探讨ASCs移植的安全性.方法 对正常脂肪组织和脂肪瘤组织进行切片染色,分离培养ASCs和LMSCs,观察细胞形态;MTS比色法检测细胞活性并绘制细胞生长曲线;流式细胞仪测定细胞周期及表面分子表达;QRT-PCR检测高迁移率族蛋白2（HMGA2）表达水平;免疫组织化学染色法鉴定端粒酶逆转录酶（hTERT）的表达.结果 正常脂肪组织和脂肪瘤组织切片差异明显;ASCs细胞形态一致性好,而LMSCs具有不均质性;MTS活性测定ASCs增殖活性要远低于LMSCs细胞（P=0.000）;流式细胞仪检测结果显示ASCs与LMSCs在干细胞标志CD29、CD44、CD105上表达类似,而在肿瘤干细胞标志CD133表达上,ASCs（5.35%）要远低于LMSCs（26.87%）;细胞周期显示ASCs的增殖能力低于LMSCs;定量PR-PCR显示ASCs中HMGA2平均aQ值为1,远低于在LMSCs中的表达（1.79）,两者差异具有统计学意义（P〈0.01）;免疫细胞化学结果：hTERT在ASCs和LMSCs中的累计吸光度A值分别为1 379.597±498.617和3 328.108±902.856,面积分别为132 390.27±35 568.945和238 000.53±49 264.289,平均吸光度A值分别为0.009±0.003和0.014±0.003,ASCs中hTERT表达远低于LMSCs.结论 体外培养的ASCs生物学特性与LMSCs存在显著差异,未发现其恶性转、化为肿瘤干细胞的证据.Objective To compared the biological characteristics of adipose-derived stem cells and lipoma-derived mesenchymal stem cells（ LMSCs） in vitro, so as to assess the safety of adipose-derived stem cells（ASCs） for transplantation. Methods Regular slice and stain of adipose and lipoma tissue were performed. ASCs and LMSCs were isolation from the two tissues by enzymatic digestion, and the appearance of the cultured cells was observed. The cell viability was evaluated with MTS chromatometry and cell growth curve was generated. Flow cytometry was performed for cell cycle analysis and the expression of the cell surface marker profiles. QRT-PCR was used to detect the expression of tumor-specific gene （the high-mobility group AT-hook 2, HMGA2） , and immunocytochemistry was used to detect the expression of telomerase. Results Marked difference was observed in histologic sections of adipose tissue and lipoma tissue. ASCs showed a good consistent in cell morphology while LMSCs not. ASCs showed a significant lower proliferation capacity than LMSCs by MTS chromatometry （ P =0. 000）. The expression of CD29, CD44, CD105 was similar in ASCs and LMSCs, while the level of CD133 was significantly lower in ASCs（5. 35% ） than in LMSCs（26. 87% ）. The expression of HMGA2 was lower in ASCs（ RQ = 1 ） than in LMSCs（ RQ = 1. 79） by qRT-PCR,it has statistically difference between them（P 〈0. 01） ; And in ASCs and LMSCs, the integrated optical intensity（ IA） values of hTERT expression are 1 379. 597 ±498. 617 and 3 328. 108 ±902.856, size（area） are 132 390. 27 ± 35 568.945 and 238 000.53 ±49 264.289, density （ mean） are 0. 009 ±0. 003 and 0. 014 ±0. 003 , revealed the expression of hTERT also shown a significant lower level in ASCs than in LMSCs by immunocytochemistry. Conclusions It indicates significant difference between ASCs and LMSCs in the biological characteristics in vitro. There is no evidence of malignant transformation of ASCs.

关 键 词：脂肪组织来源干细胞 脂肪瘤间充质干细胞 成瘤性 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]