Summary: iPS cells or induced pluripotent stem cells are stem cell-like cells derived from reprogrammed somatic cells. iPS cells reprogramming is achieved by simultaneously expressing 2-4 ES cell specific transcriptional factors including Oct4, Sox2, Myc and Klf4/Lin28 using viral vectors (lentivirus or retrovirus). The reprogramming process takes 15-30 days. First, somatic cells such as fibroblast cells are infected with a mixture of viral particles expressing the 2-4 stem cell factors. The infected cells are then seeded on mouse embryonic fibroblast (MEF) feeder cells and cultured in ES cell medium. After 10-15 days, ES like colonies of cells will appear, can be picked and expanded. Reprogramming can also be done without using MEF feeder cells.
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Protocols
- Generation of Human induced Pluripotent Stem Cells
(Center for iPS Cell Research and Application, Kyoto University)
Protocol for human iPS cell generation in a practical way focusing on problems the authors previously encountered. A unique step is the introduction of mouse solute carrier family 7 (cationic amino acid transporter, y+ system), member 1 (Slc7a1) gene encoding the ecotropic retrovirus receptor into human cells. This step enables ecotropic retroviruses to transduce human cells
http://www.cira.kyoto-u.ac.jp/e/research/images/pr...
Added: Fri Nov 06 2009, Hits: 3713, Reviews: 0 Write review - Reprogramming iPS Cells with Lentivirus (NIH Stem Cell Information)
http://stemcells.nih.gov/research/nihresearch/scun...
Added: Mon May 20 2013, Hits: 970, Reviews: 0 Write review Cached -
Basic iPS MEF-Based cell culture
(WiCell Research Institute)
This booklet contains MEF-based iPS cell culture protocols.
https://www.wicell.org/index.php?option=com_docman...
Added: Wed Jan 28 2009, Hits: 3323, Reviews: 0 Write review -
DNA Extraction for Stem Cells
(WiCell Research Institute)
https://www.wicell.org/index.php?option=com_docman...
Added: Fri Feb 20 2009, Hits: 1896, Reviews: 0 Write review -
Feeder-Dependent Culture of hESCs and hiPSCs
(NIH Stem Cell Information)
http://stemcells.nih.gov/research/nihresearch/scun...
Added: Mon May 20 2013, Hits: 711, Reviews: 0 Write review Cached -
Generating iPS Cells from MEFS through Forced Expression of Sox-2, Oct-4, c-Myc, and Klf4
(JoVE)
Demonstrates the procedure for the generation of inducible lentiviruses that express the four transcription factors and show how to infect MEFs with these viruses in order to produce iPS cells.
http://www.jove.com/index/Details.stp?ID=734
Added: Fri Nov 06 2009, Hits: 2250, Reviews: 0 Write review Cached -
Improved Method for Generation of Mouse iPS Cells
(Center for iPS Cell Research and Application, Kyoto University)
Describe a method for the induction of pluripotency by gene transfection into mouse fibroblasts
http://www.cira.kyoto-u.ac.jp/e/research/images/pr...
Added: Fri Nov 06 2009, Hits: 2193, Reviews: 0 Write review -
Induced Pluripotent Stem (iPS) Cell Culture
(WiCell Research Institute)
Protocols for culturing established iPS cells without using feeder cells.
https://www.wicell.org/index.php?option=com_docman...
Added: Fri Feb 20 2009, Hits: 3108, Reviews: 0 Write review -
iPS Cell Media & Reagents
(Thomson Lab, University of Wisconsin)
http://ink.primate.wisc.edu/~thomson/protocol.html...
Added: Wed Jan 28 2009, Hits: 2395, Reviews: 0 Write review Cached -
Reprogramming somatic cells using cell extracts
(Collas Lab, University of Oslo, Institute of Basic Medical Sciences)
An in vitro approach for directly turning one somatic cell type into another. The system relies on the transient uptake of regulatory components from a nuclear and cytoplasmic extract derived from a “target cell” by the nucleus of a reversibly permeabilized cell (referred to as the “donor cell”). Reprogramming extracts are prepared by sonication of a target cell type and sedimentation of the coarse material.
http://www.collaslab.com/UserFiles/File/Nuclear%20...
Added: Wed Jan 28 2009, Hits: 2247, Reviews: 0 Write review -
Splitting iPS cells on MEFs
(Thomson Lab, University of Wisconsin)
http://ink.primate.wisc.edu/~thomson/protocol.html...
Added: Wed Jan 28 2009, Hits: 2124, Reviews: 0 Write review Cached