临床和实验医学杂志
臨床和實驗醫學雜誌
림상화실험의학잡지
JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE
2014年
16期
1321-1323
,共3页
宫颈癌%HeLa细胞转染%Lipofectamine2000核电转红色荧光蛋白
宮頸癌%HeLa細胞轉染%Lipofectamine2000覈電轉紅色熒光蛋白
궁경암%HeLa세포전염%Lipofectamine2000핵전전홍색형광단백
Cervical carcinoma%HeLa cell%Transfection%Lipofectamine2000%Nucleofector Kit%Red fluorescence protein
目的比较两种不同的转染方法，即Lipofectamine2000和Nucleofector Kit转染人宫颈癌Hela细胞的转染效率。方法分别选取Lipofectamine2000转染和Amax核电转（ Amax Nucleofector Kit）两种转染方法。HeLa细胞培养贴壁达到85%~90%后传代获取HeLa单细胞悬液，细胞分成2管，其中1管直接采用核电转法转染红色荧光蛋白（ DsRed）后种植于1.5 ml的细胞培养皿；另一管先种植于细胞培养皿，待贴壁24 h后采用Lipofectamine2000转染DsRed。细胞的存活率采用台盼兰进行检测。分别比较两者转染效率及转染前后细胞的存活率，进一步探讨Nucleofector Kit核电转法的高效性。结果 Lipofectamine2000的基因转染率仅为20.23%，而核电转法的转染率为90.14%，后者为前者的4.46倍。Lipo-fectamine2000转染细胞前后的存活率分别为96.34%和90.76%，而核电转组分别为95.98%和78.35%。结论细胞核电转法能高效稳定地转染人宫颈癌细胞株HeLa细胞。
目的比較兩種不同的轉染方法，即Lipofectamine2000和Nucleofector Kit轉染人宮頸癌Hela細胞的轉染效率。方法分彆選取Lipofectamine2000轉染和Amax覈電轉（ Amax Nucleofector Kit）兩種轉染方法。HeLa細胞培養貼壁達到85%~90%後傳代穫取HeLa單細胞懸液，細胞分成2管，其中1管直接採用覈電轉法轉染紅色熒光蛋白（ DsRed）後種植于1.5 ml的細胞培養皿；另一管先種植于細胞培養皿，待貼壁24 h後採用Lipofectamine2000轉染DsRed。細胞的存活率採用檯盼蘭進行檢測。分彆比較兩者轉染效率及轉染前後細胞的存活率，進一步探討Nucleofector Kit覈電轉法的高效性。結果 Lipofectamine2000的基因轉染率僅為20.23%，而覈電轉法的轉染率為90.14%，後者為前者的4.46倍。Lipo-fectamine2000轉染細胞前後的存活率分彆為96.34%和90.76%，而覈電轉組分彆為95.98%和78.35%。結論細胞覈電轉法能高效穩定地轉染人宮頸癌細胞株HeLa細胞。
목적비교량충불동적전염방법，즉Lipofectamine2000화Nucleofector Kit전염인궁경암Hela세포적전염효솔。방법분별선취Lipofectamine2000전염화Amax핵전전（ Amax Nucleofector Kit）량충전염방법。HeLa세포배양첩벽체도85%~90%후전대획취HeLa단세포현액，세포분성2관，기중1관직접채용핵전전법전염홍색형광단백（ DsRed）후충식우1.5 ml적세포배양명；령일관선충식우세포배양명，대첩벽24 h후채용Lipofectamine2000전염DsRed。세포적존활솔채용태반란진행검측。분별비교량자전염효솔급전염전후세포적존활솔，진일보탐토Nucleofector Kit핵전전법적고효성。결과 Lipofectamine2000적기인전염솔부위20.23%，이핵전전법적전염솔위90.14%，후자위전자적4.46배。Lipo-fectamine2000전염세포전후적존활솔분별위96.34%화90.76%，이핵전전조분별위95.98%화78.35%。결론세포핵전전법능고효은정지전염인궁경암세포주HeLa세포。
Objective Human cervical carcinoma cell line HeLa is one of the most common used cells in the present research. In order to get the high transfection,we compared two different transfected methods,which is lipofectamine 2000 and Amax nucleofector kit. Methods We selected two kinds of transfected methods,which were the Lipofectamine2000 transfection and Amax nuclear transfer(Amax nucleofector kit). The Hela cells were passaged for single cell suspension when the cells adherent culture reached 85% ~90%. The cells were divided into two bottles, and the one was transfected directly with red fluorescent protein(DsRed),and planted in 1. 5 ml cell culture dish. Another one was transfected the DsRed with lipofectamine2000 after cell plating for 24 h. The transfected efficiency rate,the cell survival rate of the two kinds of methods were compared. Meanwhile,further study were explored about the high efficiency of Nucleofector Kit method. Results The efficiency rate of lipofactamine 2000 was only 20. 23%,however,it can reach up to 90. 14% with nucleofector kit,which was 4. 46 times than the former. The survival rate before and after transfection was 96. 34% and 90. 76%,whereas they were 95. 98% and 78. 35% respectively in nucleofector transfected group. Conclusion The method of using the nucleofector kit can transfected high efficiency for Hela cells.
