大连医科大学学报
大連醫科大學學報
대련의과대학학보
JOURNAL OF DALIAN MEDICAL UNIVERSITY
2014年
2期
132-136
,共5页
李深%白玉萌%蓝晓艳%秦华民%王苏平
李深%白玉萌%藍曉豔%秦華民%王囌平
리심%백옥맹%람효염%진화민%왕소평
细胞培养%大鼠%骨髓间充质干细胞%碱性成纤维细胞生长因子
細胞培養%大鼠%骨髓間充質榦細胞%堿性成纖維細胞生長因子
세포배양%대서%골수간충질간세포%감성성섬유세포생장인자
cell culture%rat%bone marrow mesenchymal stem cells%basic fibroblast growth factor
目的：比较4种培养条件下大鼠骨髓间充质干细胞（BMSCs）的生长与增殖性差异。方法采用密度梯度离心法提取大鼠BMSCs ，分别在DMEM-LG培养基、DMEM/F12培养基、DMEM-LG＋10 ng/mL碱性成纤维细胞生长因子（ bFGF）与DMEM/F12＋10 ng/mL bFGF培养条件下贴壁培养，观察各代细胞的形态特征，绘制第4代细胞的生长曲线，利用流式细胞仪对各培养条件下的BMSCs进行细胞表面标志CD45、CD29和CD90的检测。结果不同培养条件下的 BMSCs 生长速度不同。加入 bFGF 培养条件下细胞增殖速度较单用 DMEM -LG 或DMEM/F12培养条件下更快（P＜0．05）。 DMEM-LG＋10 ng/mL bFGF的培养效果最佳，细胞密度高，生长状态良好。4种条件下培养的细胞均阳性表达CD29及CD90，阴性表达CD45。结论短期培养过程中加入bFGF不引起BMSCs分化，DMEM-LG＋10 ng/mL bFGF是较理想的BMSCs培养条件。
目的：比較4種培養條件下大鼠骨髓間充質榦細胞（BMSCs）的生長與增殖性差異。方法採用密度梯度離心法提取大鼠BMSCs ，分彆在DMEM-LG培養基、DMEM/F12培養基、DMEM-LG＋10 ng/mL堿性成纖維細胞生長因子（ bFGF）與DMEM/F12＋10 ng/mL bFGF培養條件下貼壁培養，觀察各代細胞的形態特徵，繪製第4代細胞的生長麯線，利用流式細胞儀對各培養條件下的BMSCs進行細胞錶麵標誌CD45、CD29和CD90的檢測。結果不同培養條件下的 BMSCs 生長速度不同。加入 bFGF 培養條件下細胞增殖速度較單用 DMEM -LG 或DMEM/F12培養條件下更快（P＜0．05）。 DMEM-LG＋10 ng/mL bFGF的培養效果最佳，細胞密度高，生長狀態良好。4種條件下培養的細胞均暘性錶達CD29及CD90，陰性錶達CD45。結論短期培養過程中加入bFGF不引起BMSCs分化，DMEM-LG＋10 ng/mL bFGF是較理想的BMSCs培養條件。
목적：비교4충배양조건하대서골수간충질간세포（BMSCs）적생장여증식성차이。방법채용밀도제도리심법제취대서BMSCs ，분별재DMEM-LG배양기、DMEM/F12배양기、DMEM-LG＋10 ng/mL감성성섬유세포생장인자（ bFGF）여DMEM/F12＋10 ng/mL bFGF배양조건하첩벽배양，관찰각대세포적형태특정，회제제4대세포적생장곡선，이용류식세포의대각배양조건하적BMSCs진행세포표면표지CD45、CD29화CD90적검측。결과불동배양조건하적 BMSCs 생장속도불동。가입 bFGF 배양조건하세포증식속도교단용 DMEM -LG 혹DMEM/F12배양조건하경쾌（P＜0．05）。 DMEM-LG＋10 ng/mL bFGF적배양효과최가，세포밀도고，생장상태량호。4충조건하배양적세포균양성표체CD29급CD90，음성표체CD45。결론단기배양과정중가입bFGF불인기BMSCs분화，DMEM-LG＋10 ng/mL bFGF시교이상적BMSCs배양조건。
Objective To study the growth and proliferation differences of bone marrow mesenchymal stem cells (BMSCs) in 4 different culture conditions .Methods BMSCs were isolated from the bone marrow of rats by density gradient centrifuga-tion, and cultured in DMEM -LG, DMEM/F12, DMEM-LG+10 ng/mL basic fibroblast growth factor ( bFGF) and DMEM/F12+10 ng/mL bFGF respectively.The morphologies of each generation of BMSCs , the growth curve of the 4th generations and the expressions of CD 45, CD29 and CD90 of BMSCs in each culture conditions were detected .Results The growths of BMSCs were different among 4 culture conditions .There was statistical difference in the proliferation of BMSCs in the bFGF supplement groups in contrast to DMEM -LG and DMEM/F12 (P<0.05).DMEM-LG+10 ng/mL bFGF was the best culture condition as it achieved the highest cell density and best growth state of BMSCs .Flow cytometry analy-sis showed that BMSCs cultured in 4 different conditions highly expressed CD 29 and CD90, but did not express CD45. Conclusion Short-term supplement of bFGF did not lead to differentiation of BMSCs .DMEM-LG+10 ng/mL bFGF is an ideal culture condition for BMSCs .
