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l-Proline and GelMA hydrogel complex：An efficient antifreeze system for cell cryopreservation
Xin Li1, Yukun Cao1, Chenxi Liu1
1Institute of Biothermal Science & Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China; Shanghai Co-innovation Center for Energy Therapy of Tumors, Shanghai, 200093, China; Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shanghai, 200093, China.
This study introduces a new cell cryopreservation method using l-proline (L-Pro) and gelatin methacryloyl (GelMA) hydrogels. This biocompatible system significantly improves cell survival rates, offering a promising alternative to toxic cryoprotectant agents.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Materials Science
Background:
- Cryopreservation is vital for biomedical applications but limited by toxic cryoprotectant agents (CPAs).
- Dimethyl sulfoxide (DMSO) is a common CPA but exhibits toxicity.
- Developing safer, effective CPAs is crucial for preserving biological sample quality and function.
Purpose of the Study:
- To develop a novel, biocompatible cryopreservation system.
- To evaluate the efficacy of l-proline (L-Pro) combined with gelatin methacryloyl (GelMA) hydrogels as a CPA alternative.
- To optimize cryopreservation protocols for enhanced cell viability.
Main Methods:
- Developed a novel preservation system using l-proline (L-Pro) and gelatin methacryloyl (GelMA) hydrogels.
- Co-cultured model 3T3 cells with L-Pro and GelMA to assess biocompatibility.
- Optimized cryopreservation procedures, including cooling rates and pre-incubation times.
- Utilized Differential Scanning Calorimetry (DSC) to analyze thermal properties.
Main Results:
- L-Pro and GelMA demonstrated excellent biocompatibility compared to DMSO.
- Rapid cooling was identified as the optimal cooling procedure.
- A combination of 6% L-Pro + 7% GelMA achieved an 80% survival rate after 3 hours of co-incubation.
- The L-Pro/GelMA mixture lowered the freezing point to -4.2°C and increased unfrozen water content to 20%.
Conclusions:
- The L-Pro and GelMA hydrogel system offers a novel, biocompatible strategy for cell cryopreservation.
- This approach overcomes the limitations associated with traditional toxic CPAs.
- This study presents a new avenue for improving the quality and functionality of cryopreserved biological samples.
