Reprogramming Adipose Tissue-Derived Mesenchymal Stem Cells into Pluripotent Stem Cells by a Mutant Adeno-Associated Viral Vector

被引:7
作者
Chen, Mong-Jen [1 ]
Lu, Yuanqing [1 ]
Hamazaki, Takashi [2 ,3 ]
Tsai, Hsin-Yin [1 ]
Erger, Kirsten [4 ,5 ]
Conlon, Thomas [4 ,5 ]
Elshikha, Ahmed S. [1 ]
Li, Hong [1 ]
Srivastava, Arun [4 ,5 ]
Yao, Chunli [1 ,6 ]
Brantly, Mark [7 ]
Chiodo, Vince [3 ,5 ]
Hauswirth, William [3 ,5 ]
Terada, Naohiro [2 ,8 ]
Song, Sihong [1 ,5 ,8 ]
机构
[1] Univ Florida, Dept Pharmaceut, Gainesville, FL 32610 USA
[2] Univ Florida, Dept Pathol, Gainesville, FL 32610 USA
[3] Univ Florida, Dept Mol Genet & Microbiol, Gainesville, FL 32610 USA
[4] Univ Florida, Dept Pediat, Gainesville, FL 32610 USA
[5] Univ Florida, Powell Gene Therapy Ctr, Gainesville, FL 32610 USA
[6] Jilin Univ, Hosp 2, Dept Dermatol, Changchun 130041, Peoples R China
[7] Univ Florida, Dept Med, Gainesville, FL 32610 USA
[8] Univ Florida, Ctr Cellular Reprogramming, Gainesville, FL 32610 USA
关键词
LEBERS CONGENITAL AMAUROSIS; HUMAN FIBROBLASTS; GENE-THERAPY; HUMAN ALPHA-1-ANTITRYPSIN; EXOGENOUS SOX2; VIRUS VECTORS; SOMATIC-CELLS; GENERATION; INDUCTION; MUSCLE;
D O I
10.1089/hgtb.2013.011
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Induced pluripotent stem (iPS) cells have great potential for personalized regenerative medicine. Although several different methods for generating iPS cells have been reported, improvement of safety and efficiency is imperative. In this study, we tested the feasibility of using a triple tyrosine mutant AAV2 (Y444 + 500 + 730F) vector, designated AAV2.3m, to generate iPS cells. We developed a polycistronic rAAV2.3m vector expressing three reprogramming factors, Klf4, Oct4, and Sox2, and then used this vector to infect mouse adipose-derived mesenchymal stem cells (AT-MSCs) to induce the generation of iPS cells. We demonstrated that (1) the triple tyrosine mutant AAV2 vector is able to reprogram mouse adult adipose tissue-derived stem cells into the pluripotent state. Those rAAV2.3m-derived iPS (rAAV2.3m-iPS) cells express endogenous pluripotency-associated genes including Oct4, Sox2, and SSEA-1, and form teratomas containing multiple tissues in vivo; (2) c-myc, an oncogene, is dispensable in rAAV2.3m-mediated cellular reprogramming; and (3) transgene expression is undetectable after reprogramming, whereas vector DNA is detectable, indicating that transgenes are silenced. These results indicated the rAAV vector may have some advantages in generating iPS cells.
引用
收藏
页码:72 / 82
页数:11
相关论文
共 54 条
[1]   Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells [J].
Aasen, Trond ;
Izpisua Belmonte, Juan Carlos .
NATURE PROTOCOLS, 2010, 5 (02) :371-382
[2]  
Aoki T, 2010, TISSUE ENG PT A, V16, P2197, DOI [10.1089/ten.tea.2009.0747, 10.1089/ten.TEA.2009.0747]
[3]   A purified population of multipotent cardiovascular progenitors derived from primate pluripotent stem cells engrafts in postmyocardial infarcted nonhuman primates [J].
Blin, Guillaume ;
Nury, David ;
Stefanovic, Sonia ;
Neri, Tui ;
Guillevic, Oriane ;
Brinon, Benjamin ;
Bellamy, Valerie ;
Ruecker-Martin, Catherine ;
Barbry, Pascal ;
Bel, Alain ;
Bruneval, Patrick ;
Cowan, Chad ;
Pouly, Julia ;
Mitalipov, Shoukhrat ;
Gouadon, Elodie ;
Binder, Patrice ;
Hagege, Albert ;
Desnos, Michel ;
Renaud, Jean-Francois ;
Menasche, Philippe ;
Puceat, Michel .
JOURNAL OF CLINICAL INVESTIGATION, 2010, 120 (04) :1125-1139
[4]   Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells [J].
Brambrink, Tobias ;
Foreman, Ruth ;
Welstead, G. Grant ;
Lengner, Christopher J. ;
Wernig, Marius ;
Suh, Heikyung ;
Jaenisch, Rudolf .
CELL STEM CELL, 2008, 2 (02) :151-159
[5]   E-Cadherin-Mediated Cell-Cell Contact Is Critical for Induced Pluripotent Stem Cell Generation [J].
Chen, Taotao ;
Yuan, Detian ;
Wei, Bin ;
Jiang, Jing ;
Kang, Jiuhong ;
Ling, Kun ;
Gu, Yijun ;
Li, Jinsong ;
Xiao, Lei ;
Pei, Gang .
STEM CELLS, 2010, 28 (08) :1315-1325
[6]   Induction of pluripotent stem cells from adult somatic cells by protein-based reprogramming without genetic manipulation [J].
Cho, Hyun-Jai ;
Lee, Choon-Soo ;
Kwon, Yoo-Wook ;
Paek, Jae Seung ;
Lee, Sun-Hee ;
Hur, Jin ;
Lee, Eun Ju ;
Roh, Tae-Young ;
Chu, In-Sun ;
Leem, Sun-Hee ;
Kim, Youngsoo ;
Kang, Hyun-Jae ;
Park, Young-Bae ;
Kim, Hyo-Soo .
BLOOD, 2010, 116 (03) :386-395
[7]   Gene Therapy Using Adeno-Associated Virus Vectors [J].
Daya, Shyam ;
Berns, Kenneth I. .
CLINICAL MICROBIOLOGY REVIEWS, 2008, 21 (04) :583-593
[8]   Circular intermediates of recombinant adeno-associated virus have defined structural characteristics responsible for long-term episomal persistence in muscle tissue [J].
Duan, DS ;
Sharma, P ;
Yang, JS ;
Yue, YP ;
Dudus, L ;
Zhang, YL ;
Fisher, KJ ;
Engelhardt, JF .
JOURNAL OF VIROLOGY, 1998, 72 (11) :8568-8577
[9]   Reprogramming of Neural Progenitor Cells into Induced Pluripotent Stem Cells in the Absence of Exogenous Sox2 Expression [J].
Eminli, Sarah ;
Utikal, Jochen ;
Arnold, Katrin ;
Jaenisch, Rudolf ;
Hochedlinger, Konrad .
STEM CELLS, 2008, 26 (10) :2467-2474
[10]   Vitamin C Enhances the Generation of Mouse and Human Induced Pluripotent Stem Cells [J].
Esteban, Miguel Angel ;
Wang, Tao ;
Qin, Baoming ;
Yang, Jiayin ;
Qin, Dajiang ;
Cai, Jinglei ;
Li, Wen ;
Weng, Zhihui ;
Chen, Jiekai ;
Ni, Su ;
Chen, Keshi ;
Li, Yuan ;
Liu, Xiaopeng ;
Xu, Jianyong ;
Zhang, Shiqiang ;
Li, Feng ;
He, Wenzhi ;
Labuda, Krystyna ;
Song, Yancheng ;
Peterbauer, Anja ;
Wolbank, Susanne ;
Redl, Heinz ;
Zhong, Mei ;
Cai, Daozhang ;
Zeng, Lingwen ;
Pei, Duanqing .
CELL STEM CELL, 2010, 6 (01) :71-79