山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2014年
10期
4-6
,共3页
侯吉锋%姚星宇%张国华%赵鹏伟%杨丽敏
侯吉鋒%姚星宇%張國華%趙鵬偉%楊麗敏
후길봉%요성우%장국화%조붕위%양려민
间充质干细胞,脐带%神经巢蛋白%实时荧光定量聚合酶链反应
間充質榦細胞,臍帶%神經巢蛋白%實時熒光定量聚閤酶鏈反應
간충질간세포,제대%신경소단백%실시형광정량취합매련반응
mesenchymal stem cells,umbilical cord%nestin%real-time fluorescence quantitative polymerase chain reaction
目的:从基因角度求证脐带间充质干细胞(MSCs)神经巢蛋白nestin mRNA表达。方法取新鲜脐带组织行MSCs分离、培养及鉴定。采用胰酶消化法计数MSCs,流式细胞仪检测细胞活性,提取原代、一代、二代细胞的mR-NA,采用实时荧光定量PCR法检测nestin mRNA表达。结果脐带中分离出的细胞贴壁生长状况较好,其中大部分细胞呈梭形生长,传代后细胞与原代细胞生长形态相似,生长加快;个别细胞呈神经样生长。 MSCs呈nestin mRNA阳性表达,其表达量在传代后有明显减少;与nestin蛋白在神经前体细胞向神经细胞分化时的变化一致。结论脐带MSCs存在nestin mRNA表达;传代后nestin mRNA表达量虽减小,但仍可作为备选种子细胞进行移植。
目的:從基因角度求證臍帶間充質榦細胞(MSCs)神經巢蛋白nestin mRNA錶達。方法取新鮮臍帶組織行MSCs分離、培養及鑒定。採用胰酶消化法計數MSCs,流式細胞儀檢測細胞活性,提取原代、一代、二代細胞的mR-NA,採用實時熒光定量PCR法檢測nestin mRNA錶達。結果臍帶中分離齣的細胞貼壁生長狀況較好,其中大部分細胞呈梭形生長,傳代後細胞與原代細胞生長形態相似,生長加快;箇彆細胞呈神經樣生長。 MSCs呈nestin mRNA暘性錶達,其錶達量在傳代後有明顯減少;與nestin蛋白在神經前體細胞嚮神經細胞分化時的變化一緻。結論臍帶MSCs存在nestin mRNA錶達;傳代後nestin mRNA錶達量雖減小,但仍可作為備選種子細胞進行移植。
목적:종기인각도구증제대간충질간세포(MSCs)신경소단백nestin mRNA표체。방법취신선제대조직행MSCs분리、배양급감정。채용이매소화법계수MSCs,류식세포의검측세포활성,제취원대、일대、이대세포적mR-NA,채용실시형광정량PCR법검측nestin mRNA표체。결과제대중분리출적세포첩벽생장상황교호,기중대부분세포정사형생장,전대후세포여원대세포생장형태상사,생장가쾌;개별세포정신경양생장。 MSCs정nestin mRNA양성표체,기표체량재전대후유명현감소;여nestin단백재신경전체세포향신경세포분화시적변화일치。결론제대MSCs존재nestin mRNA표체;전대후nestin mRNA표체량수감소,단잉가작위비선충자세포진행이식。
Objective To observe the expression of nestin nestin mRNA in umbilical cord mesenchymal stem cells (MSCs) from the perspective of gene .Methods The fresh umbilical cord tissues underwent MSCc separation and then were cultured and purified .Cells from the umbilical cord were counted by enzymatic digestion , and the activity was detec-ted by flow cytometry .The primary, first-generation and second-generation cellular mRNA were extracted to detect the ex-pression of nestin mRNA by real-time PCR.Results The cells isolated from umbilical cord grew in good condition , in which a large part of the cells grew in fusiform , cells after passage had the similar morphology with primary cells and grew faster.There were individual cells which were inneural-like growth.Nestin mRNA expression was found in umbilical cord MSCs, and the expression level after the passage was significantly decreased , meanwhile, the nestin protein changes were consistent with that of differentiation from the neural precursor cells into neural cells .Conclusion The nestin mRNA ex-pression was found in the MSCs tissue genes .Even the expression level of nestin mRNA decreases after passage , it still can be used as an alternative seed cells for transplantation .