BAC Transgenesis in Human Embryonic Stem Cells as a Novel Tool to Define the Human Neural Lineage

被引:55
作者
Placantonakis, Dimitris G. [1 ,7 ]
Tomishima, Mark J. [2 ,3 ]
Lafaille, Fabien [2 ]
Desbordes, Sabrina C. [2 ]
Jia, Fan [8 ]
Socci, Nicholas D. [4 ]
Viale, Agnes [5 ,6 ]
Lee, Hyojin [2 ]
Harrison, Neil [8 ]
Tabar, Viviane [1 ]
Studer, Lorenz [1 ,2 ]
机构
[1] Sloan Kettering Inst Canc Res, Dept Neurosurg, New York, NY USA
[2] Sloan Kettering Inst Canc Res, Dev Biol Program, New York, NY USA
[3] Sloan Kettering Inst Canc Res, SKI Stem Cell Res Facil, New York, NY USA
[4] Sloan Kettering Inst Canc Res, Computat Biol Ctr, New York, NY USA
[5] Sloan Kettering Inst Canc Res, Program Mol Biol, New York, NY USA
[6] Sloan Kettering Inst Canc Res, Genom Core Lab, New York, NY USA
[7] Cornell Univ, New York Presbyterian Hosp, Weill Med Coll, Dept Neurol Surg, New York, NY 10021 USA
[8] Cornell Univ, New York Presbyterian Hosp, Weill Med Coll, Dept Pharmacol & Anesthesiol, New York, NY 10021 USA
关键词
BACTERIAL ARTIFICIAL CHROMOSOMES; CENTRAL-NERVOUS-SYSTEM; DIRECTED DIFFERENTIATION; DOPAMINERGIC-NEURONS; MOTOR-NEURONS; EXPRESSION; TRANSPLANTATION; DERIVATION; NUCLEOFECTION; MOTONEURONS;
D O I
10.1634/stemcells.2008-0884
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Human embryonic stem cells (hESCs) have enormous potential for applications in basic biology and regenerative medicine. However, harnessing the potential of hESCs toward generating homogeneous populations of specialized cells remains challenging. Here we describe a novel technology for the genetic identification of defined hESC-derived neural cell types using bacterial artificial chromosome (BAC) transgenesis. We generated hESC lines stably expressing Hes5::GFP, Dll1::GFP, and HB9::GFP BACs that yield green fluorescent protein (GFP)(+) neural stem cells, neuroblasts, and motor neurons, respectively. Faithful reporter expression was confirmed by cell fate analysis and appropriate transgene regulation. Prospective isolation of HB9::GFP(+) cells yielded purified human motor neurons with proper marker expression and electrophysiological activity. Global mRNA and microRNA analyses of Hes5::GFP(+) and HB9::GFP(+) populations revealed highly specific expression signatures, suggesting that BAC transgenesis will be a powerful tool for establishing expression libraries that define the human neural lineage and for accessing defined cell types in applications of human disease. STEM CELLS 2009; 27: 521 - 532
引用
收藏
页码:521 / 532
页数:12
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