白膜层细胞来源的富血小板血浆的制备及其对间充质干细胞增殖特性的影响  

Preparation of Platelet-Rich Plasma from the White Slurry and Its Effect on MSC Proliferation

作　　者：王克涛[1] 李众利[1] 朱恒[2] 秦园园[3] 杨以萌 李冀[1] 吕汝举[1] 刘元林[2] 毛宁[2] 张毅[2] 

机构地区：[1]中国人民解放军总医院骨科,北京100853 [2]军事医学科学院基础医学研究所,北京100850 [3]中国人民解放军总医院输血科,北京100853

出　　处：《中国实验血液学杂志》2017年第1期164-170,共7页Journal of Experimental Hematology

基　　金：军事医学创新专项(13CXZ030);国家自然科学基金面上项目(81371945;81572159;31370916)

摘　　要：目的:从成分血分离过程中废弃的白膜层细胞中分离富血小板血浆(platelet-rich plasma,PRP),为实验研究和临床应用提供大量获得PRP的途径。方法:比较白膜层细胞来源的PRP与全血来源的PRP的制备步骤,电镜观察不同PRP的形态特点,采用血细胞分析仪比较其成分构成,流式细胞术检测PRP中血小板特异标记CD41a和CD42b的表达情况,CCK-8法检测PRP对间充质干细胞增殖的影响。结果:白膜层细胞来源的PRP与全血来源的PRP具有相似的形态,接近的成份构成,表达较高水平的CD41a和CD42b;更为重要的是白膜层细胞来源的PRP能够显著促进间充质干细胞增殖。结论:白膜层细胞可以用来制备PRP,其具有和全血来源PRP相似的生物学特性,且收获量更多。本研究探索了一种可以大量获取PRP的可能途径,这为PRP相关的实验研究提供有利的支持。Objective：To isolate platelet-rich plasma（PRP） from the white slurry（WS）,a depleted fraction of the clinical blood supply,so as to provide an easier method to harvest PRP for related studies and clinical use.Methods：The protocols preparing PRP from whole blood and WS were compared.The morphological characteristics of the different PRPs were observed under transmission electron microscope;the expression of the platelet markers CD41 a and CD42 b were detected by the flow cytometry.Moreover,the ingredients of the PRPs were measured by using cytoanalyzer.for detecting the physiological function of the PRP,the harvested PRP were added to MSC culture and the cell proliferation was detected by using CCK-8 method.Results：a large amount of PRP from WS was easier harvested,the WSderived PRP shared similar morphological characteristics and ingredients as compared with whole blood-derived PRP.Importantly,the WS-derived PRP exhibited a higher expression of CD41 a and CD42 b than that of traditional PRP,which indicate that the WS is a promising reservoir for PRP.Conclusion：The WS can be used to prepare PRP,and the novel PRP share similar biological characteristics as traditional PRP prepared from whole blood.The present study provides an easier and economical method to harvest PRP and this findings may be helpful for PRP related studies.

关 键 词：富血小板血浆 白膜层 间充质干细胞 细胞增殖 

分 类 号：R457.14[医药卫生—治疗学] R331.143[医药卫生—临床医学]