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Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification
Published on: November 18, 2011
Cryopreservation of mouse embryoid bodies
Hinako Ichikawa1, Heesung No, Shunsuke Takei
1Department of Anatomy and Organ Technology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan. hinako-i@sch.md.shinshu-u.ac.jp
Cryobiology
|April 20, 2007
Summary
Cryopreservation of embryoid bodies (EBs) using the Program Freezer method resulted in higher survival and differentiation rates compared to the BICELL method. This finding is crucial for advancing regenerative therapies with stem cells.
Area of Science:
- Stem Cell Biology
- Cryobiology
- Regenerative Medicine
Background:
- Embryonic stem (ES) cells are vital for regenerative therapies.
- Cryopreservation is essential for establishing ES cell resources.
- Embryoid bodies (EBs) are used to assess ES cell multipotency.
Purpose of the Study:
- To evaluate the efficacy of cryopreservation methods for mouse ES cell-derived EBs.
- To compare the survival, structure, gene expression, and multipotency of EBs cryopreserved using BICELL and Program Freezer methods.
Main Methods:
- Mouse ES cell-derived EBs were cryopreserved using BICELL and Program Freezer systems.
- Thawed EBs were cultured for one week.
- Survival adhesion rates, cell structure, gene expression (RT-PCR), and multipotency (teratoma formation) were assessed.
Main Results:
- The Program Freezer method yielded significantly higher survival adhesion rates (55-69%) compared to BICELL (30-38%).
- While some cell damage occurred, EBs from the Program Freezer showed better structural integrity.
- RT-PCR confirmed expression of genes from all three embryonic germ layers in thawed EBs.
- Transplanted EBs formed teratomas, demonstrating multipotency.
Conclusions:
- The Program Freezer is a superior method for cryopreserving EBs, ensuring higher survival and multipotency.
- Cryopreserved EBs retain their ability to differentiate into derivatives of all three embryonic germ layers.
- This research supports the use of cryopreserved EBs for regenerative medicine applications.
