Macrocycle molecule-based cryoprotectants for ice recrystallization inhibition and cell cryopreservation

Area of Science:

• Biochemistry
• Materials Science
• Cell Biology

Background:

• Cryoprotective agents (CPAs) are crucial for preserving cells during cryopreservation.
• Developing effective and scalable CPAs remains a significant challenge in cell biology and regenerative medicine.

Purpose of the Study:

• To develop novel cyclodextrin-based cryoprotectants for improved cell preservation.
• To evaluate the efficacy of alpha-trans-methylated cyclodextrin (α-TMCD) in enhancing cell viability and cryoprotective effects.
• To investigate the synergistic mechanisms of α-TMCD in combination with dimethyl sulfoxide (DMSO).

Main Methods:

• Synthesis and characterization of cyclodextrin-based cryoprotectants, specifically α-TMCD.
• Assessment of cell viability for GES-1, human umbilical cord mesenchymal stem cells (hucMSCs), and A549 cells using α-TMCD.
• Microscopic imaging to quantify the percentage of viable hucMSCs.
• Evaluation of a cryoprotective agent (CPA) formulation based on α-TMCD and DMSO.

Main Results:

• α-TMCD demonstrated significant enhancement in cell viabilities: 19.97 ± 0.78% for GES-1, 13.93 ± 4.46% for hucMSCs, and 19.10 ± 0.95% for A549 cells.
• Light microscope imaging revealed a 103.2% enhanced hucMSC cell number percentage with α-TMCD.
• An α-TMCD-DMSO based CPA showed enhanced cryoprotective effects, attributed to DMSO-enhanced cell penetration and synergistic interaction with α-TMCD.

Conclusions:

• α-TMCD is a promising cyclodextrin-based cryoprotectant with potential for large-scale production.
• The synergistic combination of α-TMCD and DMSO offers enhanced cryoprotective capabilities.
• This study highlights the potential of α-TMCD in the discovery of novel macrocycle-molecule-based cryoprotectants for various cell types.