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Updated: Jul 19, 2026
11:38
RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells
Published on: November 26, 2018
In vitro reprogramming of nuclei and cells
Anne-Mari Håkelien1, Thomas Küntziger, Kristine G Gaustad
1Institute of Medical Biochemistry, University of Oslo, Oslo, Norway.
Methods in Molecular Biology (Clifton, N.J.)
|September 22, 2006
Summary
Scientists developed a new cell reprogramming method. This technique uses cellular extracts to convert somatic cells into other types, aiding therapeutic cell production.
Area of Science:
- Cell Biology
- Biotechnology
- Immunology
Background:
- Direct somatic cell reprogramming offers therapeutic potential for generating replacement cells.
- Novel strategies are crucial for advancing cell conversion technologies.
- Existing methods often require complex genetic manipulations.
Purpose of the Study:
- To present a novel in vitro method for functional cell reprogramming using cellular extracts.
- To demonstrate the feasibility of converting somatic cells into a different cell type without genetic alteration.
- To establish a system for studying the mechanisms of nuclear reprogramming.
Main Methods:
- Reversible permeabilization of 293T fibroblasts.
- Incubation with a nuclear and cytoplasmic extract from T-cells.
- Cell resealing and subsequent culture for reprogramming assessment.
Main Results:
- Successful nuclear uptake and assembly of transcription factors observed.
- Induction of chromatin remodeling complex activity and altered chromatin composition.
- Activation of T-cell-specific genes and surface molecule expression confirmed.
Conclusions:
- The described method enables functional reprogramming of somatic cells using cell extracts.
- This technique provides a powerful in vitro tool for investigating nuclear reprogramming processes.
- The findings support the potential of extract-based reprogramming for therapeutic applications.
Related Concept Videos
01:14Introduction to Nuclear Reprogramming
Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
01:24Methods of Nuclear Reprogramming
Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for injury repair.
01:29Somatic 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...
01:32Chromatin 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...
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...
01:13Induced 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...