来源于乳鼠与成年大鼠的脂肪源性干细胞增殖特性比较  

Proliferation characteristics of adipose-derived stem cells from neonatal suckling rats and adult ones

作　　者：杜谋选[1] 李鹏[1] 蔡颖谦[1] 邹雨汐[1] 唐艳萍[1] 秦玲莎[1] 姜晓丹[1] 

机构地区：[1]南方医科大学珠江医院神经外科,广东神经外科研究所,广东省脑功能修复与再生重点实验室,广州510280

出　　处：《中华神经医学杂志》2012年第8期770-774,共5页Chinese Journal of Neuromedicine

基　　金：广东省科技计划重点项目（2008A030201019）;广东省高校优秀青年创新人才培育项目（C1031083）;国家临床重点专科建设项目

摘　　要：目的比较来源于出生5dSD乳鼠和成年SD大鼠的脂肪源性干细胞（ADSCs）在同等条件下进行培养时增殖情况的差异。方法分别分离乳鼠和成年大鼠皮下脂肪组织并使用I型胶原酶消化法获取ADSCs，进行形态学观察。应用流式细胞仪检测ADSCs表面标记物CD45、CD29、CD90的表达情况，并在同等条件下接种相同数量的两种第2代ADSCs，培养4d后行细胞计数，比较ADSCs数量的差异。于96孔板上对相同数量的两种第2代ADSCs进行体外培养，应用CCK-8和alamarblue试剂盒连续7d对细胞增殖情况进行观察，酶标仪检测吸光度值．并绘制增殖曲线。结果流式细胞仪检测到来源于乳鼠和成年大鼠的ADSCs表面标志物均显示干细胞特性：CD45表达均呈阴性，CD29表达率分别是98．04％和93．17％，CD90表达率分别是94．92％和93.3％。在接种数量、培养条件和培养时间相同情况下，细胞计数结果显示，来源于乳鼠的ADSCs增殖数量[（8．87±0．13）×10^5]明显高于成年大鼠[（4．51±0．36）×10^5]。来源于乳鼠的ADSCs的吸光度值在第6、7天（CCK-8检测）和第2-7天（alamar blue检测）均明显高于来源于成年大鼠的ADSCs．比较差异均有统计学意义（P〈0．05）。结论来源于乳鼠的ADSCs比来源于成年大鼠的ADSCs增殖能力更强。Objective To investigate the proliferative differences of adipose-derived stem cells （ADSCs） from neonatal suckling SD rats （5-d-old） and adult ones under the same culture condition. Methods ADSCs were isolated from the subcutaneous adipose tissues of neonatal suckling SD rats and adult ones, and then, type I collagenase digestion was employed to obtain the ADSCs; the morphology of these cells was detected. The expressions of such cell surface markers as CD45, CD29 and CD90 were observed. The number of ADSCs on the 4~ d of culture under the same condition and with the same planted density was compared between the neonate and adult rats. In vitro culture of the second generation of ADSCs was performed in the 96-well plates, and CCK-8 and alamar blue kit were employed to compare and quantitate the proliferative differences; optical density was observed by microplate reader. Results The ADSCs from neonatal SD rats and adult ones expressed the stem cell biomarkers： the expression of CD45 was negative, and that of CD29 was 98.04% and 93.17%, respectively, and that of CD90 was 94.92% and 93.3%, respectively, for neonate SD rat and adult ones. The cell counting results indicated that the number of ADSCs from neonatal rats （[8.87±0.13]×10^5 cells）was larger than that of adult ones （[4.51±0.36]×10^5 cells） after being cultured under the same condition and at the same planted density. The optical density value of ADSCs in neonatal rats was significantly higher than that in adult ones on the 6^th and 7^th d of culturing detected by CCK-8 kit and on the 2^nd-7th d of culturing by alamar blue assay. Conclusion The proliferative ability of ADSCs from neonatal rats is greater than that of adult ones.

关 键 词：脂肪源性干细胞 细胞增殖 乳鼠 成年鼠 细胞培养 细胞表面标记物 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]