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Maintenance of the ES Cell State

Maintenance of the ES Cell State

The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...

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Updated: Jun 7, 2026

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

Published on: July 24, 2014

Functional assays for human embryonic stem cell pluripotency.

Michael D O'Connor¹, Melanie D Kardel, Connie J Eaves

¹Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada.

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

|November 3, 2010

Summary

Standardized methods are crucial for human embryonic stem cells (hESCs). The alkaline phosphatase colony-forming cell (AP(+)-CFC) assay is the most sensitive method for quantifying undifferentiated hESCs.

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Area of Science:

Stem cell biology
Regenerative medicine

Background:

Human embryonic stem cells (hESCs) hold significant therapeutic potential.
Standardized methods are needed to assess and modify hESC properties for biomedical advancement.

Purpose of the Study:

To highlight the importance of standardized assays for hESC research.
To introduce the alkaline phosphatase colony-forming cell (AP(+)-CFC) assay as a superior method for quantifying undifferentiated hESCs.

Main Methods:

The study emphasizes the 7-day in vitro alkaline phosphatase colony-forming cell (AP(+)-CFC) assay.
Compares AP(+)-CFC assay to phenotypic assessment, embryoid body formation, and teratoma formation in immunodeficient mice.

Main Results:

The AP(+)-CFC assay is the most sensitive and specific method for quantifying undifferentiated hESCs.
This assay is superior to phenotypic assessments for monitoring hESC culture quality.

Conclusions:

The AP(+)-CFC assay is a valuable tool for monitoring hESC quality and evaluating changes in pluripotent cell numbers.
Standardized methods like AP(+)-CFC assay are essential for realizing the full potential of hESCs in research and therapy.