Cryopreservation of Adherent Human Pluripotent Stem Cells via a Novel Dish Culture Preservation Method.

BACKGROUND

Human induced pluripotent stem cells (hiPSCs) are valuable resources for cell therapy and drug discovery. Cryopreservation is a key technique used to realize these applications, which require a large number of cells. However, standard protocols for the preservation of the adherent cells involve multiple complicated steps, which can lead to technical difficulties.

OBJECTIVE

To develop a more efficient method for cryopreservation of adherent cells using culture dishes.

MATERIALS AND METHODS

Ice-seeding treatment was employed to avoid intercellular freezing, and rapid warming used to improve cell viability.

RESULTS

The immediate survival rate after thawing was 48%. The recovery period of cells cryopreserved by the dish culture method was shortened upon subsequent passage culture, and the time for re-cultured cells to reach the appropriate confluency was reduced by two days.

CONCLUSION

The hiPSCs can be successfully cryopreserved in culture dishes with improved viability and faster recovery. The optimization of the ice-seeding temperature and cooling rate increased the survival rate.