解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2009年
4期
680-684
,共5页
孙海梅%季凤清%王屹%王丹妮%赵春礼%杨慧
孫海梅%季鳳清%王屹%王丹妮%趙春禮%楊慧
손해매%계봉청%왕흘%왕단니%조춘례%양혜
脐血间充质干细胞%淋巴细胞%增殖%羧基荧光素乙酰乙酸%四甲基偶唑盐比色法
臍血間充質榦細胞%淋巴細胞%增殖%羧基熒光素乙酰乙痠%四甲基偶唑鹽比色法
제혈간충질간세포%림파세포%증식%최기형광소을선을산%사갑기우서염비색법
Umbilical cord blood mesenchymal stem cell%Lymphocyte%Proliferation%Carboxyfluorescein didcetate%MTT assay
目的 比较羧基荧光素乙酰乙酸(CFSE)与四甲基偶氮唑盐(MTT)比色法检测人脐血间充质干细胞(CB-MSCs)和神经分化的CB-MSCs对大鼠脾淋巴细胞(LCs)刺激作用的敏感程度. 方法 分别制备刺激细胞和LCs,将刺激细胞分为4组:1. CB-MSCs组;2. Dif-CB-HSCs组:脐血间充质干细胞培养7d后维甲酸(RA)处理4d诱导其神经分化细胞;3. SH-SY5Y组:人源SH-SY5Y细胞(人神经母细胞瘤细胞系)作为阳性对照组;4. Auto-LC组:自体大鼠LCs作为阴性对照组.分别将上述各组刺激细胞与LCs进行混合培养,分别通过酶标仪(MTT法),流式细胞术(CFSE法)检测LCs增殖情况( n =3). 结果 CFSE法和MTT法检测结果 均显示,CB-MSCs组和Dif-CB-MSCs组刺激LCs增殖明显弱于阳性对照组(SH-SY5Y细胞组),但CFSE法检测结果 显示,CB-MSCs组和Dif-CB-MSCs组细胞刺激LCs增殖百分率高于阴性对照Auto-LC组,MTT法检测结果 显示,CB-MSCs组和Dif-CB-MSCs组的吸光度( A )值稍低于阴性对照Auto-LC组. 结论 CFSE法比MTT法能更加客观地反映淋巴细胞增殖情况,在实际应用中更具有优势.
目的 比較羧基熒光素乙酰乙痠(CFSE)與四甲基偶氮唑鹽(MTT)比色法檢測人臍血間充質榦細胞(CB-MSCs)和神經分化的CB-MSCs對大鼠脾淋巴細胞(LCs)刺激作用的敏感程度. 方法 分彆製備刺激細胞和LCs,將刺激細胞分為4組:1. CB-MSCs組;2. Dif-CB-HSCs組:臍血間充質榦細胞培養7d後維甲痠(RA)處理4d誘導其神經分化細胞;3. SH-SY5Y組:人源SH-SY5Y細胞(人神經母細胞瘤細胞繫)作為暘性對照組;4. Auto-LC組:自體大鼠LCs作為陰性對照組.分彆將上述各組刺激細胞與LCs進行混閤培養,分彆通過酶標儀(MTT法),流式細胞術(CFSE法)檢測LCs增殖情況( n =3). 結果 CFSE法和MTT法檢測結果 均顯示,CB-MSCs組和Dif-CB-MSCs組刺激LCs增殖明顯弱于暘性對照組(SH-SY5Y細胞組),但CFSE法檢測結果 顯示,CB-MSCs組和Dif-CB-MSCs組細胞刺激LCs增殖百分率高于陰性對照Auto-LC組,MTT法檢測結果 顯示,CB-MSCs組和Dif-CB-MSCs組的吸光度( A )值稍低于陰性對照Auto-LC組. 結論 CFSE法比MTT法能更加客觀地反映淋巴細胞增殖情況,在實際應用中更具有優勢.
목적 비교최기형광소을선을산(CFSE)여사갑기우담서염(MTT)비색법검측인제혈간충질간세포(CB-MSCs)화신경분화적CB-MSCs대대서비림파세포(LCs)자격작용적민감정도. 방법 분별제비자격세포화LCs,장자격세포분위4조:1. CB-MSCs조;2. Dif-CB-HSCs조:제혈간충질간세포배양7d후유갑산(RA)처리4d유도기신경분화세포;3. SH-SY5Y조:인원SH-SY5Y세포(인신경모세포류세포계)작위양성대조조;4. Auto-LC조:자체대서LCs작위음성대조조.분별장상술각조자격세포여LCs진행혼합배양,분별통과매표의(MTT법),류식세포술(CFSE법)검측LCs증식정황( n =3). 결과 CFSE법화MTT법검측결과 균현시,CB-MSCs조화Dif-CB-MSCs조자격LCs증식명현약우양성대조조(SH-SY5Y세포조),단CFSE법검측결과 현시,CB-MSCs조화Dif-CB-MSCs조세포자격LCs증식백분솔고우음성대조Auto-LC조,MTT법검측결과 현시,CB-MSCs조화Dif-CB-MSCs조적흡광도( A )치초저우음성대조Auto-LC조. 결론 CFSE법비MTT법능경가객관지반영림파세포증식정황,재실제응용중경구유우세.
Objective To compare the sensitivities of the stimulation effect of human cord blood mesenchymal stem cells(CB-MSCs) and CB-MSCs of neuronal differentiation to lymphocytes(LCs) detected with carboxyfluorescein didcetate(CFSE) and MTT. Methods To prepare LCs from SD rat and divided into four group stimulating cells: 1. CB-MSCs;2. Dif-CB-HSCs;3. SH-SY5Y(positive control);4. Auto-LC(negative control).Stimulating cells were respectively Co-cultured with LCs. The proliferation of LCs was detected with MTT and CFSE ( n =3). Results CB-MSCs and Dif-CB-HSCs stimulated LCs to proliferate more weakly than positive control detected with MTT and CFSE. The quantity of proliferation of lymphocytes Co-cultured with CB-MSCs and Dif-CB-HSCs were higher than that of Auto-LC detected with CFSE. But MTT OD value of CB-MSCs and Dif-CB-HSCs was a little lower than that of Auto-LC. Statistical analysis results showed no significant difference.Conclusion CFSE can reflect proliferation status of lymphocytes better than MTT. CFSE shows more advantages in practical use.
