北华大学学报（自然科学版）
北華大學學報（自然科學版）
북화대학학보（자연과학판）
JOURNAL OF BEIHUA UNIVERSITY(NATURAL SCIENCE)
2014年
5期
610-614
,共5页
王艳双%曲晓波%李枫%谭寅凤
王豔雙%麯曉波%李楓%譚寅鳳
왕염쌍%곡효파%리풍%담인봉
CCK-8法%MTT法%BMSCs%细胞毒药物体外筛选
CCK-8法%MTT法%BMSCs%細胞毒藥物體外篩選
CCK-8법%MTT법%BMSCs%세포독약물체외사선
CCK-8%MTT%bone marrow stromal cells( BMSCs)%in vitro screening of cytotoxic drugs
目的：探讨CCK-8法和MTT法在骨髓基质干细胞( Bone marrow stromal cells,BMSCs)细胞毒实验中的最佳实验条件.方法取对数生长期的P3大鼠BMSCs,制成不同浓度的细胞悬液,于96孔培养板中培养24 h,分别用CCK-8法、MTT法检测其吸光度( A)值,根据细胞浓度与A值关系曲线确定两种方法各自的最适细胞浓度；取最适浓度的细胞于0.5,1,2,3,4和5 h 6个不同时间点,分别于450,490,595 nm处测定吸光度A值,确定两种方法最佳的检测时间和最适检测波长；采用CCK-8和MTT法检测5个浓度的鹿茸Ⅰ型胶原对BMSCs的增殖的影响.结果 CCK-8法测得的A值与细胞数目之间的线性相关性优于MTT法；CCK-8法、MTT法最佳检测波长分别为450,490 nm,最佳检测时间分别为1,4 h；在检测鹿茸Ⅰ型胶原蛋白对BMSCs的增殖的影响时,CCK-8法测得数据的线性相关性优于MTT法,并且数据偏差较MTT法小.结论 CCK-8法与MTT法检测结果相比线性相关性好,灵敏度高,准确性好.在BMSCs细胞毒检测中CCK-8法优于MTT法.
目的：探討CCK-8法和MTT法在骨髓基質榦細胞( Bone marrow stromal cells,BMSCs)細胞毒實驗中的最佳實驗條件.方法取對數生長期的P3大鼠BMSCs,製成不同濃度的細胞懸液,于96孔培養闆中培養24 h,分彆用CCK-8法、MTT法檢測其吸光度( A)值,根據細胞濃度與A值關繫麯線確定兩種方法各自的最適細胞濃度；取最適濃度的細胞于0.5,1,2,3,4和5 h 6箇不同時間點,分彆于450,490,595 nm處測定吸光度A值,確定兩種方法最佳的檢測時間和最適檢測波長；採用CCK-8和MTT法檢測5箇濃度的鹿茸Ⅰ型膠原對BMSCs的增殖的影響.結果 CCK-8法測得的A值與細胞數目之間的線性相關性優于MTT法；CCK-8法、MTT法最佳檢測波長分彆為450,490 nm,最佳檢測時間分彆為1,4 h；在檢測鹿茸Ⅰ型膠原蛋白對BMSCs的增殖的影響時,CCK-8法測得數據的線性相關性優于MTT法,併且數據偏差較MTT法小.結論 CCK-8法與MTT法檢測結果相比線性相關性好,靈敏度高,準確性好.在BMSCs細胞毒檢測中CCK-8法優于MTT法.
목적：탐토CCK-8법화MTT법재골수기질간세포( Bone marrow stromal cells,BMSCs)세포독실험중적최가실험조건.방법취대수생장기적P3대서BMSCs,제성불동농도적세포현액,우96공배양판중배양24 h,분별용CCK-8법、MTT법검측기흡광도( A)치,근거세포농도여A치관계곡선학정량충방법각자적최괄세포농도；취최괄농도적세포우0.5,1,2,3,4화5 h 6개불동시간점,분별우450,490,595 nm처측정흡광도A치,학정량충방법최가적검측시간화최괄검측파장；채용CCK-8화MTT법검측5개농도적록용Ⅰ형효원대BMSCs적증식적영향.결과 CCK-8법측득적A치여세포수목지간적선성상관성우우MTT법；CCK-8법、MTT법최가검측파장분별위450,490 nm,최가검측시간분별위1,4 h；재검측록용Ⅰ형효원단백대BMSCs적증식적영향시,CCK-8법측득수거적선성상관성우우MTT법,병차수거편차교MTT법소.결론 CCK-8법여MTT법검측결과상비선성상관성호,령민도고,준학성호.재BMSCs세포독검측중CCK-8법우우MTT법.
Objective To investigate the optimal experiment conditions for MTT and CCK-8 method in cytotoxicity tests of bone marrow stromal cells ( BMSCs ) . Methods The P3 rat bone marrow stromal cells in lograrithm growth stages were prepared to cell suspensions in different cell concentrations, the BMSCs were planted in 96-well plates to be cultured for 24h, CCK-8 and MTT methods were applied to test A value, respectively. The appropriate cell concentration ranges were determined according to the curves of cell number vs A;A values of the BMSCs in the appropriate cell concentrations at 450 ,490 ,595 nm were measured at the different time points,namely,0. 5h,1h,2h,3h,4h and 5h,respectively,to determine the optimal test time and wavelength of CCK-8 and MTT methods. CCK-8 and MTT methods were used to detect the influence of five velvet antler type I collagen at the five concentrations on the proliferation of rat BMSCs. Results The linearity of cell number vs A values detected by CCK-8 was better than that by MTT;The best test wavelength of CCK-8 and MTT method was 450 and 490 nm,respectively,and the best test time was 1 and 4 h,respectively;the linearity of measured data by CCK-8 was better than that by MTT,and the data deviation was smaller than that by MTT,when the influence of velvet antler type I collagen on the proliferation of rat BMSCs. Conclusion The linearity of CCK-8 between A and cell viability was better than that of MTT, and CCK-8 assay was better than MTT assay in the detection sensitivity and accuracy. CCK-8 method was better than MTT method in the cytotoxicity test of BMSCs.
