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Related Experiment Video

Updated: Jul 2, 2026

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.
09:38

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.

Published on: October 12, 2006

Passaging HuES human embryonic stem cell-lines with trypsin.

Erin Trish1, John Dimos, Kevin Eggan

  • 1Dept of Molecular and Cellular Biology, Harvard University, USA. etrish@mcb.harvard.edu

Journal of Visualized Experiments : Jove
|August 16, 2008
PubMed
Summary

This video demonstrates routine passaging of HuES human embryonic stem cells using trypsin. Proper technique is crucial for maintaining healthy, undifferentiated, and karyotypically normal stem cell cultures.

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Area of Science:

  • Cell Biology
  • Stem Cell Research

Background:

  • Human embryonic stem cells (HuES) are valuable research tools but prone to karyotypic abnormalities during cell culture.
  • Maintaining genetic stability is critical for reliable stem cell applications.

Purpose of the Study:

  • To demonstrate a standardized protocol for passaging HuES cell lines.
  • To emphasize methods that preserve karyotypic normality and undifferentiation.

Main Methods:

  • Washing cultures with PBS, applying 0.05% Trypsin-EDTA for short durations.
  • Gently dislodging cells, diluting with HuES media, and collecting via centrifugation.
  • Resuspending cells in fresh media at a 1:10 to 1:20 split ratio and re-plating on feeder cells.

Main Results:

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Rapid, Cost-Efficient, Enzyme-Free Passaging of Human Pluripotent Stem Cells on Feeder Cells by Ethylenediaminetetraacetic Acid-Mediated Dis-Adhesion

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Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping
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Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping

Published on: September 4, 2009

Related Experiment Videos

Last Updated: Jul 2, 2026

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.
09:38

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.

Published on: October 12, 2006

Rapid, Cost-Efficient, Enzyme-Free Passaging of Human Pluripotent Stem Cells on Feeder Cells by Ethylenediaminetetraacetic Acid-Mediated Dis-Adhesion
07:21

Rapid, Cost-Efficient, Enzyme-Free Passaging of Human Pluripotent Stem Cells on Feeder Cells by Ethylenediaminetetraacetic Acid-Mediated Dis-Adhesion

Published on: July 7, 2023

Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping
10:42

Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping

Published on: September 4, 2009

  • Successful subculturing of HuES cells using the described trypsinization method.
  • Avoidance of single-cell suspension to minimize risk of karyotypic abnormalities.

Conclusions:

  • The demonstrated trypsinization protocol effectively supports the routine passaging of HuES cells.
  • Careful adherence to the protocol, particularly regarding trypsin contact time and cell suspension, is essential for maintaining stem cell quality.