CAJ | 학술논문

目的对内皮祖细胞(EPCs)检测中常用的密度梯度离心法和两种裂解红细胞的方法进行比较,探索对血液中EPCs进行以鉴定数量、含量为目的的最简便、经济、准确的实验方法.方法取新西兰白兔动脉血存储于含肝素的真空采血管中,分别采用淋巴细胞分离液、流式细胞溶血素和红细胞裂解液对血液进行预处理.所获细胞采用抗小鼠藻蓝蛋白(APC)荧光素标记的CD3(CD3-APC)、抗小鼠藻红蛋白(PE)荧光素标记的CD45(CD45-PE)、抗小鼠异硫氰酸荧光素(FITC)标记的CD133 Prominin-1[CD133-(Prominin-1)-FITC]进行标记后通过流式细胞仪检测早期EPCs含量.结果 3种方法检测早期EPCs含量差异无统计学意义(P＞0.05),其中流式细胞溶血素法所需的血液、处理液、试管和仪器最少,方法最简便,操作中影响实验结果的因素最少,有利于重复处理和测试.结论流式细胞溶血素法是检测外周血EPCs预处理方法中最容易实施的一种方法.
목적 대내피조세포(EPCs)검측중상용적밀도제도리심법화량충렬해홍세포적방법진행비교,탐색대혈액중EPCs진행이감정수량、함량위목적적최간편、경제、준학적실험방법.방법 취신서란백토동맥혈존저우함간소적진공채혈관중,분별채용림파세포분리액、류식세포용혈소화홍세포렬해액대혈액진행예처리.소획세포채용항소서조람단백(APC)형광소표기적CD3(CD3-APC)、항소서조홍단백(PE)형광소표기적CD45(CD45-PE)、항소서이류청산형광소(FITC)표기적CD133 Prominin-1[CD133-(Prominin-1)-FITC]진행표기후통과류식세포의검측조기EPCs함량.결과 3충방법검측조기EPCs함량차이무통계학의의(P＞0.05),기중류식세포용혈소법소수적혈액、처리액、시관화의기최소,방법최간편,조작중영향실험결과적인소최소,유리우중복처리화측시.결론 류식세포용혈소법시검측외주혈EPCs예처리방법중최용역실시적일충방법.
Objective In order to find out the most convenient,economic,accurate pretreatment methods for early endothelial progenitor cells (EPCs) detection from peripheral blood,this article compared the detection result of density gradient centrifugation method and two kinds of red cells lysis methods.Methods Arterial blood of New Zealand rabbits stored in vacuum heparin containing tubes was pretreated by 3 kinds of method respectively.The first pretreated method used lymphocyte separation liquid density gradient centrifugation,the second method used flow cytometry cell hemolysin and the third method used red cells lysis solution.After that,Anti-Mouse CD3-allophycocyanin (APC),Anti-Mouse CD45-phycoerythrin (PE),Anti-Mouse CD133 (Prominin-1)-fluorescein isothiocyanate (FITC) were labeled to detect early EPCs.Results There was no significant different of percentage of early endothelial progenitor cells pretreated by 3 kinds of method (P ＞ 0.05).Of the three methods,flow cytometry cell hemolysin method was the most convenient method for using the least blood,processing liquid,vitro and Instrument,not easy to be effected and repeating easily.Conclusion Flow cytometry cell hemolysin method was the most convenient pretreatment method for test endothelial progenitor cells percentage from peripheral blood.