3种人类间充质干细胞体外增殖能力和成脂能力的比较  

Comparison of proliferative and adipogenetic potentials between three human mesenchymal stem cells

作　　者：李秀英[1] 白金萍[2] 李雪[3] 王轶敏[1,4] 

机构地区：[1]吉林大学中日联谊医院中心实验室,吉林长春130033 [2]吉林大学基础医学院病理解剖学教研室,吉林长春130021 [3]延边大学医学院药理学教研室,吉林延吉133002 [4]吉林省中科生物工程有限公司,吉林长春130012

出　　处：《吉林大学学报（医学版）》2014年第4期795-800,I0003,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(20130101167JC)

摘　　要：目的:从人足月胎儿的脐带组织和胎盘组织中分离、培养间充质干细胞(MSCs),观察人脐带组织MSCs(UC-MSCs)、胎盘组织MSCs(PL-MSCs)和胚胎组织MSCs(FT-MSCs)体外增殖能力和成脂分化潜能,为干细胞进一步应用于临床提供实验依据。方法:无菌条件下获取人足月胎儿脐带组织和胎盘组织,通过Ⅰ型胶原酶酶解法分离出UC-MSCs和PL-MSCs,FT-MSCs由中科生物工程有限公司提供,取第3或4代MSCs进行检测。倒置显微镜观察细胞形态;活细胞计数法检测细胞增殖能力;流式细胞术测定UC-MSCs、PL-MSCs和FT-MSCs细胞周期并鉴定细胞表面标志物的表达阳性率。取3种组织来源的第3代MSCs进行成脂诱导,诱导18d进行油红O染色,观察并比较3种不同组织来源MSCs的成脂能力。结果:从3种不同组织中均可获取梭形贴壁的MSCs,表面标记物CD44、CD73、CD90和CD105均呈阳性表达,CD14、CD34和CD45均为阴性表达;FT-MSCs增殖能力明显强于UC-MSCs和PL-MSCs(P<0.01),FT-MSCs、UC-MSCs和PL-MSCs的增殖指数(PI)分别为(36.66±1.30)%、(18.23±1.10)%和(10.03±1.20)%,3种MSCs的PI比较差异均有统计学意义(P<0.01)。3种MSCs经过成脂诱导液诱导后,油红O染色结果均为阳性,但是UC-MSCs体外成脂能力(油红O染色阳性率)明显强于FT-MSCs和PL-MSCs(P<0.01)。结论:3种不同组织来源MSCs在形态和细胞表面标记物等方面相似,FT-MSCs具有更强的增殖能力和增殖活性,UC-MSCs具有更强的体外成脂潜能。Objective To isolate and culture mesenchymal stem cells（MSCs）from human term umbilical cord and placenta,and to study the proliferative and adipogenic capacities of human fetus tissue MSCs（FT-MSCs）,umbilical cord MSCs（UC-MSCs）and placenta MSCs（PL-MSCs）,and to provide an experimental basis for its clinical use.Methods The human term umbilical cord and placenta were obtained in sterile condition.The UCMSCs and PL-MSCs were isolated by digestion using collagenaseⅠ,the FT-MSCs were provided by Zhongke Bioengineering Co.,Ltd.The 3th or 4th passages of MSCs were selected in this study.The cell morphology was observed by inverted microscope.The proliferative ability was detected by living cell number counting method.The cell cycle and the positive expression rates of the surface markers of three kinds of MSCs were detected by flow cytometry（FCM）.The 3th generation of UC-MSCs,PL-MSCs,and FT-MSCs were induced to differentiate into adipogenic cells.18dafter being induced,the differentiated cells were stained with Oil red O and the positive cells were counted.The adipogenic differentiation abilities of MSCs from the different tissues were assessed.Results The MSCs from different tissues were spindle-shaped adherent cells.The FCM results showed the surface marker of MSCs were positive for CD44,CD73,CD90,and CD105,and negative for CD14,CD34,and CD45.The FT-MSCs displayed higher proliferative ability than UC-MSCs and PL-MSCs（P〈0.01）.The proliferative index（PI）of FT-MSCs（36.66%±1.30%）was significantly higher than those of UC-MSCs（18.23%±1.10%）and PL-MSCs（10.03%±1.20%）（P〈0.01）.The lipid droplets were found after induction,and the results of Oil red O staining were positive in all kinds of MSCs.However the adipogenic capacity of UC-MSCs was significantly higher than those of PL-MSCs and FT-MSCs（P〈0.01）.Conclusion UC-MSCs,PL-MSCs and FT-MSCs have similar biological characteristics and cell surface markers.But FT-MSCs have the strongest proliferative ability and activity,UC

关 键 词：胚胎 胎盘 脐带 间充质干细胞 增殖能力 成脂诱导 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]