红细胞冻干保存前对海藻糖的负载  

Loading red blood cells with trehalose before lyophilization

作　　者：陈麟凤[1] 刘景汉[1] 欧阳锡林[1] 庄远[1] 车辑[1] 王珊[2] 

机构地区：[1]解放军总医院输血科全军临床输血中心,北京100853 [2]解放军总医院实验测试中心

出　　处：《中国输血杂志》2007年第4期272-275,共4页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金资助课题(编号30471931)

摘　　要：目的探寻红细胞负载海藻糖的有效方法,评价负载海藻糖对红细胞各项理化指标的影响。方法设实验组(负载海藻糖红细胞)和对照组(未负载海藻糖红细胞),使用硫酸-蒽酮法测定胞内海藻糖含量,检测负载后红细胞各项理化指标,通过流式细胞术检测负载后红细胞膜的完整性。结果在37℃条件下,红细胞对海藻糖的摄取随胞外海藻糖浓度的增加而增多,当海藻糖浓度为800mmol/L,水浴7h,红细胞负载海藻糖可达到有效浓度;且2组红细胞各项理化指标比较差异无统计学意义(P>0.05);流式结果显示红细胞在高渗环境中负载海藻糖后,细胞膜结合很少量Annexin-V-FITC,并且破损细胞能被有效清除。结论红细胞37℃孵育7h,胞外海藻糖浓度为800mmol/L,能有效摄取海藻糖,且保持红细胞的理化稳定性和膜结构完整性。Objective To optimize the method of loading red blood cell（RBC） with trehalose and evaluate the effect of trehalose loading on RBC before lyophilization. Methods The samples were divided into two groups： the control group （without loading trehalose）, and the test group（loading RBCs in trehalose buffer）. The amount of in- tracellular trehalose and integrality of RBC membrane were determined by both anthrone reaction and flow cytometry. Results The intracelluar trehalose concentration increased considerably with the increase of trehalose concentration in the extracelluar solution at 37℃. After the trehaose loading, the counting, MHC, MCHC, ATP level and G--6--PD activity between the two groups showed no significant difference （P〉0.05）. Flow cytometry data demonstrated that RBC incubated in hypertonic trehalose solution could combine a tiny amount of Annexin--V--FITC, which can be re- moved by washing and resuspending the RBC in an iso--osmotic medium. Conelusion Loading RBC with trehalose at 37℃ after 7--hour incubation is efficient. And minor damage to the RBC membrane is caused during the loading.

关 键 词：海藻糖 负载缓冲液 红细胞冻干保存 流式细胞术 

分 类 号：R331.141[医药卫生—人体生理学] R318.52[医药卫生—基础医学]