Trehalose to cryopreserve human pluripotent stem cells

Area of Science:

• Stem Cell Biology
• Cryobiology
• Biotechnology

Background:

• Human pluripotent stem cells (hPSCs) are crucial for research and therapies.
• Current cryopreservation methods using dimethyl sulfoxide (DMSO) can impair hPSC pluripotency and epigenetics.
• A simple, efficient, and safe cryopreservation method is needed for hPSCs.

Purpose of the Study:

• To evaluate trehalose-based cryosolutions for preserving hPSCs and neural stem cells (NSCs).
• To compare the efficacy of trehalose solutions with standard DMSO cryopreservation.
• To assess the impact of trehalose cryopreservation on cell viability, pluripotency, differentiation, genotoxicity, and endoplasmic reticulum homeostasis.

Main Methods:

• Testing trehalose-based cryosolutions with ethylene glycol (EG) or glycerol (GLY).
• Utilizing human embryonic stem cells (hESCs), induced pluripotent stem cells (hiPSCs), and long-term neuroepithelial-like stem cells (lt-NES).
• Assessing cell viability, morphology, self-renewal, pluripotency, differentiation potential, genotoxicity, and endoplasmic reticulum stress.

Main Results:

• Trehalose-based cryosolutions demonstrated acceptable cell viability and high stability compared to 10% DMSO.
• All tested cell lines (hESCs, hiPSCs, lt-NES) retained morphology, self-renewal, and pluripotency.
• Differentiation potential remained unaffected; genotoxicity varied, but endoplasmic reticulum homeostasis was preserved.

Conclusions:

• Trehalose-based cryopreservation is effective for hPSCs and lt-NES, maintaining their functional properties.
• This method offers a viable alternative to DMSO, potentially supporting cell therapy applications under GMP conditions.