中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2010年
1期
18-22
,共5页
冉玉琴%冯丽娜%徐燕%唐琪%慕晓玲
冉玉琴%馮麗娜%徐燕%唐琪%慕曉玲
염옥금%풍려나%서연%당기%모효령
间质干细胞%表型%细胞周期%转化
間質榦細胞%錶型%細胞週期%轉化
간질간세포%표형%세포주기%전화
Mesenchymal stem cells%Phenotype%Cell cycle%Transformations
目的 观察抗砷细胞模型慢性砷暴露人骨髓间充质干细胞(CAsE-hFBMSCs)的生物学特性,探讨长期低剂量砷暴露对人骨髓间充质干细胞(hFBMSCs)的影响.方法 常规条件培养hFBMSCs,48 h急性砷毒性实验MTS法检测不同剂量砷刺激条件下[0(对照)、0.25、0.50、1.00、2.00、4.00、8.00、20.00、40.00、80.00、120.00μmol/L]第2代(P2)hFBMSCs细胞生存率,根据检测结果用1.00μmol/L亚砷酸钠刺激hFBMSCs 14周,建立抗砷细胞模型CAsE-hFBMSCs,作为实验组,同时设立对照组,并采用荧光激活细胞分选术鉴定诱导前后细胞表型,流式细胞术检测第2、9、16代(P2、P9、P16)细胞周期,软琼脂克隆形成实验检测细胞是否发生恶性转化.结果 <10μmol/L时,急性砷刺激促进hFBMSCs增殖,而≥10μmol/L时,急性砷刺激则抑制细胞的生长.14周时实验组半数致死剂量(LC_(50))为(89.42±0.64)μmol/L,对照组为(52.48±0.71)μmol/L,组间比较差异有统计学意义(t=123.89,P<0.05);抗砷细胞模型CAsE-hFBMSCs的细胞表型CD34、CD45表达阴性,CD29、CD90、CD166表达阳性,与对照组比较细胞表型未见明显改变;CAsE-hFBMSCs的细胞周期变化较大,与对照组[(8.44±0.45)%、(9.14±0.14)%、(82.42±0.60)%]比较,P2实验组G2/M期[(17.72±5.47)%]和S期细胞[(25.34±3.36)%]增加,G0/G1期细胞减少[(56.96±8.83)%],P16细胞周期恢复至与对照组相近的水平:软琼脂克隆形成实验中,抗砷细胞CAsE-hFBMSCs未见克隆形成.结论 长期低剂量砷刺激对hFBMSCs生物学特性无明显影响.
目的 觀察抗砷細胞模型慢性砷暴露人骨髓間充質榦細胞(CAsE-hFBMSCs)的生物學特性,探討長期低劑量砷暴露對人骨髓間充質榦細胞(hFBMSCs)的影響.方法 常規條件培養hFBMSCs,48 h急性砷毒性實驗MTS法檢測不同劑量砷刺激條件下[0(對照)、0.25、0.50、1.00、2.00、4.00、8.00、20.00、40.00、80.00、120.00μmol/L]第2代(P2)hFBMSCs細胞生存率,根據檢測結果用1.00μmol/L亞砷痠鈉刺激hFBMSCs 14週,建立抗砷細胞模型CAsE-hFBMSCs,作為實驗組,同時設立對照組,併採用熒光激活細胞分選術鑒定誘導前後細胞錶型,流式細胞術檢測第2、9、16代(P2、P9、P16)細胞週期,軟瓊脂剋隆形成實驗檢測細胞是否髮生噁性轉化.結果 <10μmol/L時,急性砷刺激促進hFBMSCs增殖,而≥10μmol/L時,急性砷刺激則抑製細胞的生長.14週時實驗組半數緻死劑量(LC_(50))為(89.42±0.64)μmol/L,對照組為(52.48±0.71)μmol/L,組間比較差異有統計學意義(t=123.89,P<0.05);抗砷細胞模型CAsE-hFBMSCs的細胞錶型CD34、CD45錶達陰性,CD29、CD90、CD166錶達暘性,與對照組比較細胞錶型未見明顯改變;CAsE-hFBMSCs的細胞週期變化較大,與對照組[(8.44±0.45)%、(9.14±0.14)%、(82.42±0.60)%]比較,P2實驗組G2/M期[(17.72±5.47)%]和S期細胞[(25.34±3.36)%]增加,G0/G1期細胞減少[(56.96±8.83)%],P16細胞週期恢複至與對照組相近的水平:軟瓊脂剋隆形成實驗中,抗砷細胞CAsE-hFBMSCs未見剋隆形成.結論 長期低劑量砷刺激對hFBMSCs生物學特性無明顯影響.
목적 관찰항신세포모형만성신폭로인골수간충질간세포(CAsE-hFBMSCs)적생물학특성,탐토장기저제량신폭로대인골수간충질간세포(hFBMSCs)적영향.방법 상규조건배양hFBMSCs,48 h급성신독성실험MTS법검측불동제량신자격조건하[0(대조)、0.25、0.50、1.00、2.00、4.00、8.00、20.00、40.00、80.00、120.00μmol/L]제2대(P2)hFBMSCs세포생존솔,근거검측결과용1.00μmol/L아신산납자격hFBMSCs 14주,건립항신세포모형CAsE-hFBMSCs,작위실험조,동시설립대조조,병채용형광격활세포분선술감정유도전후세포표형,류식세포술검측제2、9、16대(P2、P9、P16)세포주기,연경지극륭형성실험검측세포시부발생악성전화.결과 <10μmol/L시,급성신자격촉진hFBMSCs증식,이≥10μmol/L시,급성신자격칙억제세포적생장.14주시실험조반수치사제량(LC_(50))위(89.42±0.64)μmol/L,대조조위(52.48±0.71)μmol/L,조간비교차이유통계학의의(t=123.89,P<0.05);항신세포모형CAsE-hFBMSCs적세포표형CD34、CD45표체음성,CD29、CD90、CD166표체양성,여대조조비교세포표형미견명현개변;CAsE-hFBMSCs적세포주기변화교대,여대조조[(8.44±0.45)%、(9.14±0.14)%、(82.42±0.60)%]비교,P2실험조G2/M기[(17.72±5.47)%]화S기세포[(25.34±3.36)%]증가,G0/G1기세포감소[(56.96±8.83)%],P16세포주기회복지여대조조상근적수평:연경지극륭형성실험중,항신세포CAsE-hFBMSCs미견극륭형성.결론 장기저제량신자격대hFBMSCs생물학특성무명현영향.
Objective To study the biological characteristics of arsenic resistance cell model chronic arsenic exposure human bone marrow mesenchymal stem cells (CAsE-hFBMSCs) and discuss the consequence of chronic arsenite exposure to human mesenchymal stem cells (hFBMSCs). Methods hFBMSCs cultivated under general conditions,hFBMSC cell survival rate was detected in 48 hours with arsenite toxicity test under different doses arsenic [0(control),0.25,0.50,1.00,2.00,4.00,8.00,20.00,40.00,80.00,120.00 μmol/L]of the fist 2-generation(P2). According to the test results,1.00 μmol/L sodium arsenite was chosen to stimulate hFBMSCs for 14 weeks as experimental group,simultaneous 0 μmol/L sodium arsenite as the control group. And then,the phenotype was detected by fluorescence-activated cell sorting,and the cell cycle by flow cytometry. Finally,the cell malignant transformation was detected by soft-agar assay. Results Arsenite low than 10 μmol/L promoted cell proliferation,but inhibited cell proliferation when exceeding 10 μmol/L. Half of the lethal dose (LC_(50)) in experimental and control groups were (89.42±0.64),(52.48±0.71)μmol/L. The difference between two groups was statistically significant(t = 123.89,P < 0.05). The phenotype of CAsE-hFBMSCs was CD29,CD90,CD166 positive and CD34,CD45 negative. The phenotype of CAsE-hFBMSCs was the same as the control. Comparing to control group[(8.44±0.45)%,(9.14μ0.14)%,(82.42±0.60)%],G2/M phage[(17.72±5.47)%]and S phage [(25.34±3.36)%]cell increased,G0/G1 phage[(56.96±8.83)%]cell decreased in P2 CAsE-hFBMSCs. The cell cycle became nearly the same as the control group after adaption. CAsE-hFBMSCs did not show clone formation in soft agar clone formation assay. Conclusion Long last and low level exposure to arsenite does not influence the biologic features of hFBMSCs.
