Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance

	Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance
	Luo CH(罗春花); Lv DY(吕东媛); Pan J; Long M(龙勉)
刊名	ACS APPLIED MATERIALS & INTERFACES
	2016
通讯作者邮箱	panj@cqu.edu.cn ; mlong@imech.ac.cn
卷号	8 期号:13 页码:8367-8375
关键词	dendrimer embryonic stem cells gene transfection end group size endocytosis
ISSN号	1944-8244
通讯作者	Pan, J (reprint author), Chongqing Univ, Coll Bioengn, Minist Educ, Key Lab Biorheol Sci & Technol, Chongqing 400044, Peoples R China.
产权排序	[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
中文摘要	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.
分类号	一类
类目[WOS]	Nanoscience & Nanotechnology ; Materials Science, Multidisciplinary
研究领域[WOS]	Science & Technology - Other Topics ; Materials Science
关键词[WOS]	dendrimer ; embryonic stem cells ; gene transfection ; end group ; size ; endocytosis
收录类别	SCI ; EI
原文出处	http://dx.doi.org/10.1021/acsami.6b00353
语种	英语
WOS记录号	WOS:000373748200007
内容类型	期刊论文
源URL	[http://dspace.imech.ac.cn/handle/311007/59506]
专题	力学研究所_国家微重力实验室
推荐引用方式 GB/T 7714	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.
APA	Luo CH,Lv DY,Pan J,&Long M.(2016).Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance.ACS APPLIED MATERIALS & INTERFACES,8(13),8367-8375.
MLA	Luo CH,et al."Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance".ACS APPLIED MATERIALS & INTERFACES 8.13(2016):8367-8375.