Related Experiment Video
Updated: Jun 10, 2026
08:49
Freezing and Thawing Human Embryonic Stem Cells
Published on: December 24, 2009
Thawing embryonic stem (ES) cells from a 96-well plate
Cold Spring Harbor Protocols
|July 22, 2010
Summary
This study details a crucial method for thawing frozen embryonic stem (ES) cells after genome modification. Efficiently rescuing these genetically altered cells ensures successful downstream genomic analysis and research.
Area of Science:
- Genomics
- Stem Cell Biology
- Molecular Biology
Background:
- Embryonic stem (ES) cells are commonly modified using electroporation to integrate exogenous DNA.
- Post-modification, ES cells are cultured, cryopreserved in 96-well plates, and genotyped via Southern blot or PCR.
- Effective recovery of cryopreserved ES cells is vital for subsequent research.
Purpose of the Study:
- To describe a reliable protocol for thawing cryopreserved, genetically modified embryonic stem cells.
- To optimize the recovery process for ES cells stored after genome alteration.
Main Methods:
- Cryopreservation of genetically modified ES cells in 96-well plates.
- Development and validation of a specific thawing protocol for these cells.
Main Results:
- Successful recovery of viable embryonic stem cells from cryopreserved stocks.
- Demonstration of the protocol's effectiveness for downstream applications like genotyping.
Conclusions:
- The described thawing method is essential for maintaining the integrity of genetically modified ES cells.
- This protocol facilitates the continuation of research involving genome-edited stem cell lines.
