SD大鼠外周血单个核细胞成分及干/祖细胞含量的年龄变化  

Age-related changes in the composition of peripheral blood mononuclear cells and the content of stem/progenitor cells in SD rats

作　　者：张文博 陈祖祥 刘镇 孙爱军 蔺海燕 单乐天 赵东宝 Zhang Wenbo;Chen Zuxiang;Liu Zhen;Sun Aijun;Lin Haiyan;Shan Letian;Zhao Dongbao(Department of Rheumatology and Immunology,Changhai Hospital,Shanghai 200433;Zhejiang Institute of Orthopedics&Traumatology,Hangzhou 311400,China;Department of Human Anatomy,Naval Medical University,Shanghai 200433)

机构地区：[1]海军军医大学长海医院风湿免疫科,上海200433 [2]浙江省骨伤研究所,杭州311400 [3]海军军医大学人体解剖学教研室,上海200433

出　　处：《解剖学杂志》2024年第4期295-298,313,F0002,共6页Chinese Journal of Anatomy

基　　金：国家自然科学基金(82171754,81971484)。

摘　　要：目的:研究SD大鼠外周血单个核细胞(PBMNCs)的组成成分及其随年龄的变化,为大鼠PBMNCs相关研究提供参考。方法:使用淋巴细胞分离液、不连续密度梯度离心法制备3、6、12月龄雄性SD大鼠PBMNCs,检测PBMNCs中细胞种类和含量、淋巴细胞和单核细胞含量随年龄的变化趋势;流式细胞术、免疫荧光技术检测PBMNCs干/祖细胞含量及随年龄变化趋势。结果:使用淋巴细胞分离液,不连续梯度离心法制备PBMNCs,不同细胞成分分层清晰,红细胞、粒细胞去除较完全,PBMNCs中淋巴细胞、单核细胞含量随年龄的增长下降趋势明显;PBMNCs中Nanog、OCT4、SOX2、SSEA4四种干性标志物均有表达,Nanog阳性表达量最低,SSEA4表达量最高,可达23.1%;总体趋势均为6月龄大鼠PBMNCs中干/祖细胞含量最高。结论:SD大鼠PBMNCs中干/祖细胞含量较高,动物实验治疗疾病的作用可能较人体应用的疗效好。进行SD大鼠PBMNCs移植治疗疾病的实验研究,建议选用6月龄以下大鼠为宜。Objective:To provide a reference for studies related to rat peripheral blood mononuclear cells(PBMNCs),we investigated the composition and age-related changes in PBMNCs isolated from SD rats.Methods:PBMNCs were isolated from male SD rats aged 3,6,and 12 months using lymphocyte separation medium and discontinuous density gradient centrifugation.The types and numbers of cells in PBMNCs,as well as the agerelated changes in lymphocyte and monocyte content,were analyzed.Flow cytometry and immunofluorescence were utilized to quantify the number and assess the age-related trends of stem/progenitor cells within PBMNCs.Results:The different cellular components of PBMNCs were clearly separated,and red blood cells and granulocytes were effectively removed.The number of lymphocytes and monocytes in PBMNCs decreased significantly with age.In SD rat PBMNCs,four stemness-related markers(Nanog,OCT4,SOX2,and SSEA4)were expressed.Among them,Nanog exhibited the lowest positive expression,while SSEA4 showed the highest,reaching 23.1%.Overall,the content of PBMNCs stem/progenitor cells in PBMNCs was highest in six-month-old rats.Conclusions:SD rats have a higher content of stem/progenitor cells in their PBMNCs,which suggests that the therapeutic effect of animal experiments might be better than in human applications.For experimental studies involving the transplantation of SD rat PBMNCs for disease treatment,it is recommended to use rats younger than 6 months.

关 键 词：外周血单个核细胞 干性标志物 年龄变化 SD大鼠 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学]