中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2010年
2期
125-132
,共8页
钱奕炜%高建华%鲁峰%郑旭东
錢奕煒%高建華%魯峰%鄭旭東
전혁위%고건화%로봉%정욱동
脂肪组织来源干细胞%脂肪瘤间充质干细胞%成瘤性
脂肪組織來源榦細胞%脂肪瘤間充質榦細胞%成瘤性
지방조직래원간세포%지방류간충질간세포%성류성
Adipose-derived mesenchymal stem cells%Lipoma-derived mesenchymal stem cells%Tumorigenesis
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.
目的 比較體外培養的脂肪來源榦細胞(adipose-derived stem cells,ASCs)與脂肪瘤間充質榦細胞(lipoma-derived mesenchymal stem cells,LMSCs)的生物特性,以探討ASCs移植的安全性.方法 對正常脂肪組織和脂肪瘤組織進行切片染色,分離培養ASCs和LMSCs,觀察細胞形態;MTS比色法檢測細胞活性併繪製細胞生長麯線;流式細胞儀測定細胞週期及錶麵分子錶達;QRT-PCR檢測高遷移率族蛋白2(HMGA2)錶達水平;免疫組織化學染色法鑒定耑粒酶逆轉錄酶(hTERT)的錶達.結果 正常脂肪組織和脂肪瘤組織切片差異明顯;ASCs細胞形態一緻性好,而LMSCs具有不均質性;MTS活性測定ASCs增殖活性要遠低于LMSCs細胞(P=0.000);流式細胞儀檢測結果顯示ASCs與LMSCs在榦細胞標誌CD29、CD44、CD105上錶達類似,而在腫瘤榦細胞標誌CD133錶達上,ASCs(5.35%)要遠低于LMSCs(26.87%);細胞週期顯示ASCs的增殖能力低于LMSCs;定量PR-PCR顯示ASCs中HMGA2平均aQ值為1,遠低于在LMSCs中的錶達(1.79),兩者差異具有統計學意義(P<0.01);免疫細胞化學結果:hTERT在ASCs和LMSCs中的纍計吸光度A值分彆為1 379.597±498.617和3 328.108±902.856,麵積分彆為132 390.27±35 568.945和238 000.53±49 264.289,平均吸光度A值分彆為0.009±0.003和0.014±0.003,ASCs中hTERT錶達遠低于LMSCs.結論 體外培養的ASCs生物學特性與LMSCs存在顯著差異,未髮現其噁性轉、化為腫瘤榦細胞的證據.
목적 비교체외배양적지방래원간세포(adipose-derived stem cells,ASCs)여지방류간충질간세포(lipoma-derived mesenchymal stem cells,LMSCs)적생물특성,이탐토ASCs이식적안전성.방법 대정상지방조직화지방류조직진행절편염색,분리배양ASCs화LMSCs,관찰세포형태;MTS비색법검측세포활성병회제세포생장곡선;류식세포의측정세포주기급표면분자표체;QRT-PCR검측고천이솔족단백2(HMGA2)표체수평;면역조직화학염색법감정단립매역전록매(hTERT)적표체.결과 정상지방조직화지방류조직절편차이명현;ASCs세포형태일치성호,이LMSCs구유불균질성;MTS활성측정ASCs증식활성요원저우LMSCs세포(P=0.000);류식세포의검측결과현시ASCs여LMSCs재간세포표지CD29、CD44、CD105상표체유사,이재종류간세포표지CD133표체상,ASCs(5.35%)요원저우LMSCs(26.87%);세포주기현시ASCs적증식능력저우LMSCs;정량PR-PCR현시ASCs중HMGA2평균aQ치위1,원저우재LMSCs중적표체(1.79),량자차이구유통계학의의(P<0.01);면역세포화학결과:hTERT재ASCs화LMSCs중적루계흡광도A치분별위1 379.597±498.617화3 328.108±902.856,면적분별위132 390.27±35 568.945화238 000.53±49 264.289,평균흡광도A치분별위0.009±0.003화0.014±0.003,ASCs중hTERT표체원저우LMSCs.결론 체외배양적ASCs생물학특성여LMSCs존재현저차이,미발현기악성전、화위종류간세포적증거.
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.
