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Updated: May 12, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Published on: May 29, 2014

Profiling stem cells using quantitative PCR protein assays.

David Ruff¹, Pauline T Lieu

¹Life Technologies, South San Francisco, CA, USA.

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

|April 3, 2013

Summary

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Researchers developed a quantitative PCR method to track key protein factors during induced pluripotent stem cell (iPSC) reprogramming. This technique efficiently assesses pluripotency and aids in identifying iPSC colonies for biological research.

Area of Science:

Stem cell biology
Molecular biology
Biotechnology

Background:

Induced pluripotent stem cells (iPSCs) are crucial for regenerative medicine and disease modeling.
Pluripotency markers are essential for defining stem cell status and potential.
Current methods for assessing pluripotency can be time-consuming and lack real-time kinetic data.

Purpose of the Study:

To develop and validate a real-time quantitative PCR (qPCR) methodology for assessing protein factor levels during human somatic cell reprogramming.
To apply this qPCR method for monitoring reprogramming kinetics and characterizing iPSC clones.
To demonstrate the utility of TaqMan® Protein Assays in high-throughput screening for iPSC identification.

Main Methods:

Reprogramming of human dermal fibroblasts using four transcription factors (OCT3/4, SOX2, KLF4, cMYC) delivered via viral vectors.

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Published on: October 25, 2018

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

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Last Updated: May 12, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Published on: May 29, 2014

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

Development and application of a real-time qPCR assay to quantify specific protein levels.

Analysis of protein expression kinetics over the reprogramming time course and comparison across different iPSC clones.

Main Results:

The developed qPCR methodology accurately quantifies key protein factors involved in reprogramming.
Real-time monitoring revealed the kinetics of pluripotency factor expression during the reprogramming process.
The assay facilitated the characterization of pluripotent cells and the identification of potential iPSC colonies in screening.

Conclusions:

TaqMan® Protein Assays provide a fast, simple, and sensitive method for monitoring stem cell reprogramming.
This qPCR approach generates a valuable pluripotency scorecard for assessing reprogrammed stem cells.
The methodology enhances the efficiency of iPSC generation and characterization for research applications.