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Updated: Mar 1, 2026
08:25
Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells iPSCs Using Retroviral Vector with GFP
Published on: April 3, 2012
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Fluorescent tagged episomals for stoichiometric induced pluripotent stem cell reprogramming
Christopher E Schmitt1, Blanca M Morales2,3, Ellen M H Schmitz1
1Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, 94158, USA.
Stem Cell Research & Therapy
|June 7, 2017
Summary
Modified episomal vectors now express fluorescent proteins, enabling easier identification and selection of reprogrammed cells. This advancement improves the efficiency and understanding of induced pluripotent stem cell generation.
Area of Science:
- Stem cell biology
- Molecular biology
- Epigenetics
Background:
- Non-integrating episomal vectors are crucial for induced pluripotent stem cell (iPSC) reprogramming using Yamanaka factors.
- Current methods struggle to efficiently identify cells successfully transfected with these reprogramming vectors.
- The stochastic nature of reprogramming hinders the isolation of desired cell clones.
Purpose of the Study:
- To develop modified episomal vectors for improved tracking and enrichment of reprogrammed cells.
- To assess the efficacy of these novel vectors compared to standard ones.
- To enhance the understanding of the reprogramming process and iPSC isolation.
Main Methods:
- Engineered episomal vectors to express spectrally separable fluorescent proteins.
- Tested modified vectors against original vectors for reprogramming efficiency.
- Evaluated the ability to enrich for stoichiometric ratios of reprogramming factors.
Main Results:
- Reengineered vectors facilitate cell sorting based on reprogramming factor expression.
- Demonstrated tracking of episomal expression in individual cells.
- Enabled selection of optimal reprogramming factor dosage.
Conclusions:
- Modified vectors serve as valuable tools for studying the reprogramming process.
- These vectors enhance the efficiency of induced pluripotent stem cell isolation.
- Facilitates better control and understanding of episomal vector-mediated cell reprogramming.
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