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

ABSTRACT

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.