Generation of human induced pluripotent stem cells using epigenetic regulators reveals a germ cell-like identity in partially reprogrammed colonies

PloS One
Akshi GoyalRenee A Reijo Pera

Abstract

Previous studies have shown that induced pluripotent stem cells (iPSCs) can be derived from fibroblasts by ectopic expression of four transcription factors, OCT4, SOX2, KLF4 and c-MYC using various methods. More recent studies have focused on identifying alternative approaches and factors that can be used to increase reprogramming efficiency of fibroblasts to pluripotency. Here, we use nucleofection, morpholino technologies and novel epigenetic factors, which were chosen based on their expression profile in human embryos, fibroblasts and undifferentiated/differentiated human embryonic stem cells (hESCs) and conventionally generated iPSCs, to reprogram human fibroblasts into iPSCs. By over expressing DNMT3B, AURKB, PRMT5 and/or silencing SETD7 in human fibroblasts with and without NANOG, hTERT and/or SV40 overexpression, we observed the formation of colonies resembling iPSCs that were positive for certain pluripotency markers, but exhibited minimal proliferation. More importantly, we also demonstrate that these partially-reprogrammed colonies express high levels of early to mid germ cell-specific genes regardless of the transfection approach, which suggests conversion to a germ cell-like identity is associated with early reprogr...Continue Reading

References

Nov 1, 1993·Developmental Dynamics : an Official Publication of the American Association of Anatomists·M BuehrS Darling
Feb 22, 2002·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Hidemasa GotoMasaki Inagaki
May 4, 2005·Cancer Research·Douglas C MarchionPamela N Munster
Jan 21, 2006·Development Genes and Evolution·Joyce Y TungJörg Gromoll
Jan 27, 2007·Stem Cells and Development·Kehkooi KeeRenee A Reijo Pera
Jun 8, 2007·Nature·Keisuke OkitaShinya Yamanaka
Nov 21, 2007·Current Opinion in Biotechnology·Marc Lewitzky, Shinya Yamanaka
Nov 22, 2007·Science·Junying YuJames A Thomson
Dec 7, 2007·Nature Biotechnology·Masato NakagawaShinya Yamanaka
Dec 22, 2007·Nature·Ian ChambersAustin Smith
May 30, 2008·Nature·Tarjei S MikkelsenAlexander Meissner
Jun 24, 2008·Nature Reviews. Molecular Cell Biology·Dieter EgliKevin Eggan
Oct 11, 2008·Science·Keisuke OkitaShinya Yamanaka
Oct 14, 2008·Nature Biotechnology·Danwei HuangfuDouglas A Melton
Apr 29, 2009·Cell Stem Cell·Hongyan ZhouSheng Ding
Aug 26, 2009·Cell·Jose SilvaAustin Smith
Oct 15, 2009·Nature Biotechnology·Elayne M ChanThorsten M Schlaeger
Oct 20, 2009·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Noemi FusakiMamoru Hasegawa
Jul 21, 2010·Nature·K KimG Q Daley
Jan 5, 2011·Current Biology : CB·Thorold W TheunissenJosé C R Silva
Jan 7, 2011·Cold Spring Harbor Symposia on Quantitative Biology·M R Hübner, D L Spector
Mar 19, 2011·Nature Reviews. Genetics·Kathrin Plath, William E Lowry
Apr 9, 2011·Cell Stem Cell·Tasha KalistaChristopher Thomas Scott
May 31, 2011·Cell Stem Cell·Norikatsu MiyoshiMasaki Mori

❮ Previous
Next ❯

Methods Mentioned

BETA
transfection
PCR
confocal microscopy
transfect

Software Mentioned

GENE
qBasePlus

Related Concepts

Related Feeds

Cell Adhesion Molecules in the Brain

Cell adhesion molecules found on cell surface help cells bind with other cells or the extracellular matrix to maintain structure and function. Here is the latest research on their role in the brain.

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis