The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell

DUAN Lian; et al

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The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell

更新时间：2024-01-17

- The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell
- Journal of Chongqing Medical University Vol. 34, Issue 1, Pages: 9-12(2009)
- 作者机构：
  
  重庆医科大学附属第一医院内分泌科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R587.2
- Published：2009，
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[1]段炼,周波,李启富.影响人脐静脉内皮细胞脂质体转染pReceiver-M29-PRKCB1效率因素的探讨[J].重庆医科大学学报,2009,34(01):9-12.

DUAN Lian, et al. The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell. [J]. Journal of Chongqing Medical University 34(1):9-12(2009)
[1]段炼,周波,李启富.影响人脐静脉内皮细胞脂质体转染pReceiver-M29-PRKCB1效率因素的探讨[J].重庆医科大学学报,2009,34(01):9-12. DOI：

DUAN Lian, et al. The influencing factors for efficiency of liposome-mediated transfection of pReceiver-M29-PRKCB1 into human umbilical vein endothelial cell. [J]. Journal of Chongqing Medical University 34(1):9-12(2009) DOI：

摘要

目的:探讨影响人脐静脉内皮细胞转染效率的因素

为优化外源基因在其中的表达提供实验依据。方法:选用适于转染人脐静脉内皮细胞的脂质体lipofectin

采用不同的转染条件:脂质体和DNA的用量;铺板密度;细胞与DNA-脂质体复合物孵育时间转染内皮细胞。荧光显微镜、流式细胞仪测定不同转染参数下的转染效率。结果:⑴24孔板中

铺板密度大于2×l04个/孔和孵育时间超过4h

反而使转染效率下降。⑵随质粒质量增加转染效率逐渐上升

达峰后质粒质量的增加并不能显著增高转染效率。⑶转染效率达峰前

随着脂质体用量的增加

转染效率增高;达峰后

脂质体用量的增加

转染效率反降低。当DNA（μg）和脂质体（μl）的用量比为1∶6～1∶8时

获得最高的转染效率:18.62%。结论:采用优化后的转染参数在人脐静脉内皮细胞转染中可获得较优的转染效率。

Abstract

Objective: To observe the influencing factors for efficiency of liposome-mediated transfection into human umbilical vein endothelial cell(HUVEC). Methods: Transferring HUVECs by distinct conditions

such as ratios of liposome and plasmid

densities of per well and different times of incubation

and transfection rates were observed and calculated by fluorescent microscope and flow cytometry. Results:⑴ When density of per well exceeded 2×l04 and times exceeded 4 hours

liposome-mediated transfection efficiency of endothelial cells decreased. ⑵With adding plasmid quality

transfection rates increased;while plasmid quality exceeded 1.0 μg

in condition of 2×l04 per well and liposome volume 2 μl

transfection efficiency reached the peak level. ⑶With adding liposome volume

transfection rates increased;while liposome volume was 8 μl

in condition of 2×104 per well and plasmid quality 1 μg

transfection efficiency was depressed.When ratios of liposome and plasmid were 1:6～1:8

the optimal transfection efficiency was 18.62%. Conclusion: Using optimal transfection parameter

we obtained the optimal transfection efficiency .

关键词

脂质体转染人脐静脉内皮细胞

Keywords

liposometransfectionhuman umbilical vein endothelial cell

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