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Related Concept Videos

Methods of Nuclear Reprogramming01:24

Methods 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.
Adult Stem Cells01:33

Adult Stem Cells

Stem cells are undifferentiated cells that divide and produce more stem cells or progenitor cells that differentiate into mature, specialized cell types. All the cells in the body are generated from stem cells in the early embryo, but small populations of stem cells are also present in many adult tissues including the bone marrow, brain, skin, and gut. These adult stem cells typically produce the various cell types found in that tissue—to replace cells that are damaged or to continuously renew...
Stem Cell Culture01:17

Stem Cell Culture

Stem cell research aims to find ways to use stem cells to regenerate and repair cellular damage. Over time, most adult cells undergo the wear and tear of aging and lose their ability to divide and repair themselves. Stem cells do not display a particular morphology or function. Adult stem cells, which exist as a small subset of cells in most tissues, keep dividing and can differentiate into a number of specialized cells generally formed by that tissue. These cells enable the body to renew and...
Forced Transdifferentiation01:28

Forced Transdifferentiation

Transdifferentiation, also known as lineage reprogramming, was first discovered by Selman and Kafatos in 1974 in silkmoths. They observed that the moths’ cuticle-producing cells transformed into salt-producing cells. Many such cases of natural transdifferentiation occur in organisms. In humans, pancreatic alpha cells can become beta cells. In newts, the loss of the eye’s lens causes the pigmented epithelial cells to transdifferentiate into the lens cells.
Artificial transdifferentiation occurs...
Induced Pluripotent Stem Cells01:13

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 Cells01:06

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...

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Related Experiment Video

Updated: Jul 14, 2026

Isolation of Adult Human Dermal Fibroblasts from Abdominal Skin and Generation of Induced Pluripotent Stem Cells Using a Non-Integrating Method
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A reverse transfection technology to genetically engineer adult stem cells.

Arimichi Okazaki1, Jun-Ichiro Jo, Yasuhiko Tabata

  • 1Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.

Tissue Engineering
|May 24, 2007
PubMed
Summary

A novel non-viral gene transfection method using spermine-pullulan enhanced gene expression in rat mesenchymal stem cells (MSCs). Reverse transfection on a coated substrate significantly improved expression levels and duration compared to conventional methods.

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An Optimized Mouse Embryonic Stem Cell Based Reverse Poly-Transfection Technique for Rapid Exploration of Nucleic Acid Ratios
06:04

An Optimized Mouse Embryonic Stem Cell Based Reverse Poly-Transfection Technique for Rapid Exploration of Nucleic Acid Ratios

Published on: December 8, 2023

Area of Science:

  • Biotechnology
  • Gene Delivery
  • Stem Cell Research

Background:

  • Gene transfection is crucial for stem cell research and therapy.
  • Non-viral methods offer advantages over viral vectors, including lower cytotoxicity.
  • Enhancing gene expression levels and duration in mesenchymal stem cells (MSCs) remains a challenge.

Purpose of the Study:

  • To develop and evaluate a novel non-viral gene transfection method for rat MSCs.
  • To compare the efficacy of reverse transfection with conventional transfection methods.
  • To investigate the impact of culture substrate modification and agitation on gene expression.

Main Methods:

  • Cationized pullulan (spermine-pullulan) was synthesized as a non-viral gene carrier.
  • Plasmid DNA encoding luciferase was complexed with spermine-pullulan.
  • Complexes were coated onto culture substrates with Pronectin for reverse transfection.
  • Rat MSCs were transfected using both reverse and conventional methods.
  • Reverse transfection was also performed on polyethylene terephthalate (PET) non-woven fabric using static, agitated, and stirred culture methods.

Main Results:

  • Reverse transfection significantly enhanced and prolonged gene expression compared to conventional transfection.
  • Reverse transfection allowed MSC culture in serum-containing medium, reducing cytotoxicity.
  • Agitated and stirred culture methods on PET fabric further enhanced gene expression levels and duration compared to static methods.
  • Medium circulation likely improved oxygen and nutrient supply and waste removal, enhancing gene expression.

Conclusions:

  • The developed spermine-pullulan based reverse transfection method is effective for enhancing gene expression in rat MSCs.
  • Reverse transfection offers a promising strategy for improved gene delivery in stem cell applications.
  • Optimized culture conditions, including substrate modification and medium circulation, can further boost transfection efficiency.