Manipulating the cell differentiation through lentiviral vectors.
The manipulation of cell differentiation is important to create new sources for the treatment of degenerative diseases or solve cell depletion after aggressive therapy against cancer. In this chapter, the use of a tissue-specific promoter lentiviral vector to obtain a myocardial pure lineage from murine embryonic stem cells (mES) is described in detail. Since the cardiac isoform of troponin I gene product is not expressed in skeletal or other muscle types, short mouse cardiac troponin proximal promoter is used to drive reporter genes. Cells are infected simultaneously with two lentiviral vectors, the first expressing EGFP to monitor the transduction efficiency, and the other expressing a puromycin resistance gene to select the specific cells of interest. This technical approach describes a method to obtain a pure cardiomyocyte population and can be applied to other lineages of interest.
Coppola
Valeria
V
Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome, Italy.
Galli
Cesare
C
Musumeci
Maria
M
Bonci
Désirée
D
D000818:Animals; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D011401:Promoter Regions, Genetic