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Induced Pluripotent Stem Cells

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore called induced pluripotent stem...

Induced Pluripotent Stem Cells

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic cells are...

Induced Pluripotent Stem Cells

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore called induced pluripotent stem...

Somatic to iPS Cell Reprogramming

Somatic to iPS Cell Reprogramming

Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012 for this...

Chromatin Modification in iPS Cells

Chromatin Modification in iPS Cells

Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...

Embryonic Stem Cells

Embryonic Stem Cells

Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
ES cells are grown in a culture medium where they can divide indefinitely, creating ES cell lines. Under certain conditions, ES cells can differentiate, either spontaneously into a variety of...

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Related Experiment Video

Updated: May 30, 2026

Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors

Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors

Published on: October 28, 2014

Episomal transgene expression in pluripotent stem cells.

Michele M P Lufino¹, Anna R Popplestone, Sally A Cowley

¹Molecular Neurodegeneration and Gene Therapy Research Group, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.

Methods in Molecular Biology (Clifton, N.J.)

|August 9, 2011

Summary

Herpes simplex type 1 (HSV-1) amplicon vectors efficiently deliver genes to stem cells. These large vectors allow extrachromosomal DNA retention, avoiding common issues with gene therapy vectors.

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Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells

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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

Area of Science:

Gene therapy
Stem cell biology
Virology

Background:

Herpes simplex type 1 (HSV-1) amplicon vectors are promising tools for gene delivery.
Efficient transduction of both dividing and non-dividing cells is a key advantage.
The large capacity of HSV-1 amplicons allows for the inclusion of large DNA sequences.

Purpose of the Study:

To describe the construction and use of HSV-1 amplicon vectors for stem cell transduction.
To highlight the advantages of extrachromosomal retention elements in gene delivery vectors.
To detail the packaging of amplicon vectors into HSV-1 viral particles.

Main Methods:

Construction of HSV-1 amplicon vectors with extrachromosomal retention elements.
Packaging of amplicon vectors into HSV-1 viral particles.
Transduction of stem cells using HSV-1 amplicons.

Main Results:

HSV-1 amplicon vectors can efficiently transduce both dividing and non-dividing cells.
The large size of HSV-1 amplicons permits the delivery of large genomic DNA loci with native promoters.
Extrachromosomal retention elements prevent transgene silencing and insertional mutagenesis.

Conclusions:

HSV-1 amplicon vectors are versatile and efficient tools for stem cell gene therapy.
Extrachromosomal elements offer significant advantages over integrating vectors.
The described methods facilitate the development of novel gene delivery strategies using HSV-1 amplicons.