中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2012年
8期
770-774
,共5页
杜谋选%李鹏%蔡颖谦%邹雨汐%唐艳萍%秦玲莎%姜晓丹
杜謀選%李鵬%蔡穎謙%鄒雨汐%唐豔萍%秦玲莎%薑曉丹
두모선%리붕%채영겸%추우석%당염평%진령사%강효단
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
脂肪源性榦細胞%細胞增殖%乳鼠%成年鼠%細胞培養%細胞錶麵標記物
지방원성간세포%세포증식%유서%성년서%세포배양%세포표면표기물
Adipose-derived stromal cell%Cell proliferation%Neonatal rat%Adult rat%Cell culture%Cell marker
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alamar blue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)×105]明显高于成年大鼠[(4.51±0.36)× 105].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.
目的 比較來源于齣生5dSD乳鼠和成年SD大鼠的脂肪源性榦細胞(ADSCs)在同等條件下進行培養時增殖情況的差異. 方法 分彆分離乳鼠和成年大鼠皮下脂肪組織併使用Ⅰ型膠原酶消化法穫取ADSCs,進行形態學觀察.應用流式細胞儀檢測ADSCs錶麵標記物CD45、CD29、CD90的錶達情況,併在同等條件下接種相同數量的兩種第2代ADSCs,培養4d後行細胞計數,比較ADSCs數量的差異.于96孔闆上對相同數量的兩種第2代ADSCs進行體外培養,應用CCK-8和alamar blue試劑盒連續7d對細胞增殖情況進行觀察,酶標儀檢測吸光度值,併繪製增殖麯線. 結果 流式細胞儀檢測到來源于乳鼠和成年大鼠的ADSCs錶麵標誌物均顯示榦細胞特性:CD45錶達均呈陰性,CD29錶達率分彆是98.04％和93.17％,CD90錶達率分彆是94.92％和93.3％.在接種數量、培養條件和培養時間相同情況下,細胞計數結果顯示,來源于乳鼠的ADSCs增殖數量[(8.87±0.13)×105]明顯高于成年大鼠[(4.51±0.36)× 105].來源于乳鼠的ADSCs的吸光度值在第6、7天(CCK-8檢測)和第2～7天(alamar blue檢測)均明顯高于來源于成年大鼠的ADSCs,比較差異均有統計學意義(P ＜0.05). 結論 來源于乳鼠的ADSCs比來源于成年大鼠的ADSCs增殖能力更彊.
목적 비교래원우출생5dSD유서화성년SD대서적지방원성간세포(ADSCs)재동등조건하진행배양시증식정황적차이. 방법 분별분리유서화성년대서피하지방조직병사용Ⅰ형효원매소화법획취ADSCs,진행형태학관찰.응용류식세포의검측ADSCs표면표기물CD45、CD29、CD90적표체정황,병재동등조건하접충상동수량적량충제2대ADSCs,배양4d후행세포계수,비교ADSCs수량적차이.우96공판상대상동수량적량충제2대ADSCs진행체외배양,응용CCK-8화alamar blue시제합련속7d대세포증식정황진행관찰,매표의검측흡광도치,병회제증식곡선. 결과 류식세포의검측도래원우유서화성년대서적ADSCs표면표지물균현시간세포특성:CD45표체균정음성,CD29표체솔분별시98.04％화93.17％,CD90표체솔분별시94.92％화93.3％.재접충수량、배양조건화배양시간상동정황하,세포계수결과현시,래원우유서적ADSCs증식수량[(8.87±0.13)×105]명현고우성년대서[(4.51±0.36)× 105].래원우유서적ADSCs적흡광도치재제6、7천(CCK-8검측)화제2～7천(alamar blue검측)균명현고우래원우성년대서적ADSCs,비교차이균유통계학의의(P ＜0.05). 결론 래원우유서적ADSCs비래원우성년대서적ADSCs증식능력경강.
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 4th 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]×105 cells) was larger than that of adult ones ([4.51±0.36]×105 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 6th and 7th d of culturing detected by CCK-8 kit and on the 2nd-7th d of culturing by alamar blue assay. Conclusion The proliferative ability of ADSCs from neonatal rats is greater than that of adult ones.
