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Related Experiment Video

Updated: Jul 14, 2025

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
09:51

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation

Published on: February 2, 2016

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Protocol for selecting single human pluripotent stem cells using a modified micropipetter.

Sandra P Mojica-Perez1, Kyle Stokes2, Daniel C Jaklic2

  • 1Department of Neurology, University of Michigan, Ann Arbor, MI 48109, USA; The Human Stem Cell and Gene Editing Core, University of Michigan, Ann Arbor, MI 48109, USA.

STAR Protocols
|October 7, 2023
PubMed
Summary
This summary is machine-generated.

This study presents a new protocol for isolating single human pluripotent stem cells using a repurposed micromanipulator. This method efficiently generates homogeneous clonal cultures with high survival rates, avoiding cell sorting.

Keywords:
CRISPRNeuroscienceStem Cells

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Area of Science:

  • Stem Cell Biology
  • Reproductive Technology

Background:

  • Generating homogeneous human pluripotent stem cell (hPSC) cultures is essential for research and therapeutic applications.
  • Current single-cell isolation methods can be labor-intensive or require specialized equipment like cell sorters.

Purpose of the Study:

  • To describe a novel, accessible protocol for single-cell clonal selection of hPSCs.
  • To adapt the STRIPPER Micropipetter, typically used in in vitro fertilization, for efficient stem cell isolation.

Main Methods:

  • Repurposing the STRIPPER Micropipetter for picking individual hPSCs.
  • Detailed steps for tool preparation, single-cell isolation, and colony passaging.
  • Procedures for subsequent cell culture amplification and analysis.

Main Results:

  • The protocol successfully generates homogeneous clonal hPSC cultures.
  • Achieved a survival rate exceeding 50% for isolated single cells.
  • The method does not necessitate the use of cell sorting.

Conclusions:

  • The repurposed STRIPPER Micropipetter offers a practical and effective alternative for single-cell clonal selection of hPSCs.
  • This protocol simplifies the generation of uniform stem cell populations, enhancing their utility in various research fields.