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Title: Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance
Author: Luo CH(罗春花); Lv DY(吕东媛); Pan J; Long M(龙勉)
Source: ACS APPLIED MATERIALS & INTERFACES
Issued Date: 2016
Volume: 8, Issue:13, Pages:8367-8375
Abstract: Manipulation of genes in human embryonic stem cells (hESCs) is imperative for their highly potential applications; however, the transduction efficiency remains very low. Although existing evidence revealed the type, size, and zeta potential of vector affect gene transfection efficiency in cells, the systematic study in hESCs is scarce. In this study, using poly(amidoamine) (PAMAM) dendrimers ended with amine, hydroxyl, or carboxyl as model, we tested the influences of size and surface group as well as cytotoxicity and endocytosis on hESC gene transfection. We found that in culture medium of mTeSR the particle sizes of G5, G7, G4.5COOH, and GSOH were around 5 nm and G1 had a smaller size of 3.14 nm. G5 and G7 had a slight and significant positive zeta potential, respectively, whereas G1 was slightly negative, and G4.5COOH and GSOH were significantly negative. We demonstrated that only amine-terminated dendrimers accomplished gene transfection in hESCs, which is greater than that from Lipofectamine 2000 transfection. Ten micromolar GS had the greatest efficiency and was better than 1000 mu M G1. Only a low concentration (0.5 and 1 mu M) of G7 realized gene delivery. Amine-ended dendrimers, especially with higher generations, were detrimental to the growth and pluripotent maintenance of hESCs. In contrast, similarly sized hydroxyl- and carboxyl-terminated dendrimers exerted much lower cytotoxicity, in which carboxyl-terminated dendrimer maintained pluripotency of hESCs. We also confirmed the endocytosis into and significant exocytosis from hESCs using FITC-Iabeled GS dendrimer. These results suggested that careful considerations of size, concentration, and zeta potential, particularly the identity and position of groups, as well as minimized exocytosis in the design of a vector for hESC gene delivery are necessary, which helps to better design an effective vector in hESC gene transduction.
Keyword: dendrimer ; embryonic stem cells ; gene transfection ; end group ; size ; endocytosis
Language: 英语
Indexed Type: SCI ; EI
Corresponding Author: Pan, J (reprint author), Chongqing Univ, Coll Bioengn, Minist Educ, Key Lab Biorheol Sci & Technol, Chongqing 400044, Peoples R China.
Correspondent Email: panj@cqu.edu.cn ; mlong@imech.ac.cn
DOI: 10.1021/acsami.6b00353
Related URLs: 查看原文
DOC Type: 期刊论文
WOS Subject: Nanoscience & Nanotechnology ; Materials Science, Multidisciplinary
WOS Subject Extended: Science & Technology - Other Topics ; Materials Science
WOS Keyword Plus: dendrimer ; embryonic stem cells ; gene transfection ; end group ; size ; endocytosis
WOS ID: WOS:000373748200007
ISSN: 1944-8244
Funder: We thank Dr. Ye-Guang Chen from Tsinghua University for providing us H1 hES cells upon Simple Letter Agreement between Dr. Chen's and Dr. Long's laboratories that is approved by WiCell Research Institute. This work was supported by National Natural Science Foundation of China grants 31110103918 and 31470907, Chinese Academy of Sciences Strategic Priority Research Program grants XDA01030604 and XDA04020219, National High Technology Research and Development Program of China grant 2011AA020109, and Fundamental Research Funds for the Central Universities of China grant CQKLBST-2012-002.
Rank: [Luo, Chunhua; Pan, Jun] Chongqing Univ, Coll Bioengn, Minist Educ, Key Lab Biorheol Sci & Technol, Chongqing 400044, Peoples R China; [Luo, Chunhua; Lu, Dongyuan; Long, Mian] Chinese Acad Sci, Natl Micrograv Lab, Key Lab Micrograv, Ctr Biomech & Bioengn, Beijing 100190, Peoples R China; [Luo, Chunhua; Lu, Dongyuan; Long, Mian] Chinese Acad Sci, Inst Mech, Beijing Key Lab Engn Construct & Mechanobiol, Beijing 100190, Peoples R China
Department: NML分子-细胞生物力学与空间生命科学
Classification: 一类
First Institution (Yes or Not): True
Citation statistics:
Content Type: 期刊论文
URI: http://dspace.imech.ac.cn/handle/311007/59506
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Recommended Citation:
Luo CH,Lv DY,Pan J,et al. Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance[J]. ACS APPLIED MATERIALS & INTERFACES,2016,8(13):8367-8375.
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