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Category Archives: Publications

Integrating piggyBac Transposon Transgenes into Mouse Fibroblasts Using Chemical Methods

Mouse embryonic fibroblasts can be reprogrammed to ES cell-like pluripotent stem cells by the forced expression of four transcription factors—OCT4, SOX2, KLF4, and c-MYC. The piggyBac transposon system has proven effective as a vehicle for the delivery of transgenes into fibroblasts and for successful reprogramming to induced pluripotent stem (iPS) cells. We found that FuGENE HD transfection reagent can be effective for mouse embryonic fibroblasts (MEFs) to generate induced pluripotent stem cells (iPSCs) with the piggyBac transposon transgenes. There are multitudes of cell transfection methods commercially available. Their efficiency, and thus their success, in inducing pluripotent cell types are cell-type-dependent.

  • From the Manipulating the Mouse Embryo collection, edited by Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy, and Andras Nagy.

© 2017 Cold Spring Harbor Laboratory Press

Reprogramming Mouse Fibroblasts with piggyBac Transposons

In 2006, Shinya Yamanaka and his student Kazutoshi Takahashi showed that the expression of only four specific genes is sufficient to reprogram fully differentiated somatic cells into pluripotent stem cells. These cells, termed induced pluripotent stem (iPS) cells, share many of their characteristics with embryonic stem (ES) cells. In this protocol, we describe one of the simplest ways of generating iPS cells from mouse fibroblasts. It combines an efficient transposon-mediated transfection and the tetracycline-inducible system to control the expression of the Yamanaka reprogramming factors. Continue Reading

Does Brain Tissue Regeneration Depend on Maturity of Stem Cells Used for Transplantation?

New research has shown that the success of transplanting stem cells into the brain to regenerate tissue damaged by stroke may depend on the maturity of the neuronal precursor cells used for transplantation. A study demonstrating the significant impact of human neuronal precursor cell maturity on cell survival after transplantation into stroke-injured rate brains is published in hTissue Engineering, Part A, peer-reviewed journal from Mary Ann Liebert, Inc., publishers… Continue Reading

Lentivector Iterations and Pre-Clinical Scale-Up/Toxicity Testing: Targeting Mobilized CD34+ Cells for Correction of Fabry Disease.

Fabry disease is a rare lysosomal storage disorder (LSD). We designed multiple recombinant lentivirus vectors (LVs) and tested their ability to engineer expression of human α-galactosidase A (α-gal A) in transduced Fabry patient CD34+ hematopoietic cells. We further investigated the safety and efficacy of a clinically directed vector, LV/AGA, in both ex vivo cell culture studies and animal models. …. Continue Reading

Concise Review: Wharton’s Jelly: The Rich, but Enigmatic, Source of Mesenchymal Stromal Cells.

The umbilical cord has become an increasingly used source of mesenchymal stromal cells for preclinical and, more recently, clinical studies. Despite the increased activity, several aspects of this cell population have been under-appreciated. Key issues are that consensus on the anatomical structures within the cord is lacking, and potentially different populations are identified as arising from a single source. To help address these points, we propose a histologically based nomenclature for cord structures and provide an analysis of their developmental origins and composition…. Continue Reading