File:Reprogramming MEF into ES-like cells 03.jpg

Reprogramming of genetically unmodified mouse embryonic fibroblasts into embryonic stem-like cells

Outline of the mouse embryonic fibroblasts (MEF) reprogramming protocol.

• 2.5×10⁵ MEF cells were plated in two 100 mm dishes
• were infected with Sox2, Klf4, c-Myc and Oct3/4 virus
• Cells in dish 1(blue arrow) were passage on day 8 after infection
• ES-like colonies form 4 days later
• Cells in dish 2 (red arrow) were AP staining positive on day 14 and the colonies were counted.

File:Reprogramming MEF into ES-like cells 01.jpg (B) Significant morphology changes observed on day 4 and ES-like colonies form on day 9 after MEF cells were infected with Sox2, Klf4, c-Myc and Oct3/4 virus. (C) Morphology of ES-like colonies on day 6 after passage. (D) AP positive colonies on day 14 post infection. (E) Estimated reprogramming efficiency of genetically unmodified MEF into AP positive ES-like cells 16 days post infection.

File:Reprogramming MEF into ES-like cells 02.jpg (F) ES-like cell lines were established using the nonselective approach. Representative colonies from passage 6 cells were stained with anti-Rex1, Sox2 and SSEA1 antibodies and images were acquired through a Leica confocal system.

Original file name: Cr200792f1a.jpg

Reference

<pubmed>17971807</pubmed>| Cell Research

Reprinted by permission from Macmillan Publishers Ltd

Licensee: Mark A Hill License Date: May 24, 2011 License Number: 2675351286106 Publication: Cell Research Title: Direct generation of ES-like cells from unmodified mouse embryonic fibroblasts by Oct4/Sox2/Myc/Klf4 Type Of Use: post on the internet

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