Author/Authors :
Movahedi Motlagh، Fatemeh نويسنده Department of Stem Cells and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran , , Rajaei، Bahareh نويسنده Department of Stem Cells and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, IR Iran , , Massumi، Mohammad نويسنده National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran , , Kabir-Salmani، Maryam نويسنده , , Soleimanpour-Lichaei، Hamid Reza نويسنده Department of Medical Biotechnology, Stem Cell Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran ,
Abstract :
Background
Human somatic cells are reprogrammed to induced pluripotent stem (iPS) cells when four transcription factors (Oct4, Sox2, Klf4, and Myc) are ectopically expressed in them. These iPS cells, which evade immunological rejection, are a valuable source for patient-specific cell therapy. Lentiviral systems have been proved to be powerful tools for cellular reprogramming, an example of which is the induction of iPS cells from somatic cells that requires a high transduction efficiency of lentiviruses harboring the four reprogramming factors.
Objectives
The purpose of this study was to define an optimized calcium phosphate transfection method to produce high-titer lentiviral vectors for the generation of human iPS cells.
Materials and Methods
In this study, the calcium phosphate transfection method was used to generate lentiviruses. The virus supernatant was concentrated using Amicon Ultra-4 column.
Results
This method resulted in 80% GFP-positive cells and viral preparations of 2.4 × 108 viral particles/mL.
Conclusions
This method is both cost effective and simple to adopt.
