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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Dimethyl-Sulfoxide-Free Cell Cryoprotectant Derived from Amino Acids.

Xi Chen¹, Taijie Zhan¹, Yuting Wang¹

¹Institute of Bio-thermal Science and Technology, Shanghai Co-innovation Center for Energy Therapy of Tumors, Shanghai Technical Service Platform for Cryopreservation of Biological Resources, University of Shanghai for Science and Technology, Shanghai 200093, People's Republic of China.

Langmuir : the ACS Journal of Surfaces and Colloids

|November 22, 2024

Summary

Green amino acids offer a non-toxic alternative to traditional cryoprotectants (CPAs) for cell therapy. This study developed a novel amino acid-based cryoprotectant with comparable efficacy to dimethyl sulfoxide (DMSO).

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Published on: August 5, 2011

Area of Science:

Cell biology
Biotechnology
Cryobiology

Background:

Traditional permeable cryoprotectants (CPAs) used in cell therapy cause significant cellular damage.
There is a critical need for developing green, non-toxic cryoprotectants to mitigate these issues.
Amino acids show potential as non-toxic cryoprotectants due to their metabolic roles and ability to balance osmotic pressure.

Purpose of the Study:

To investigate the efficacy of three amino acids (l-proline, l-carnitine, and betaine) as cryoprotectants.
To develop and evaluate a novel three-component mixed amino acid cryoprotectant for cell preservation.
To provide guidance for developing dimethyl sulfoxide (DMSO)-free cryopreservation methods.

Main Methods:

Single-factor experiments determined optimal concentrations for individual amino acid cryoprotectants.
Multivariate orthogonal experiments evaluated the efficacy of a three-component mixed cryoprotectant.
Evaluation criteria included thermophysical parameters, ice crystal inhibition, and post-cryopreservation cell viability.

Main Results:

The study identified optimal concentration ranges for l-proline, l-carnitine, and betaine.
The developed three-component amino acid cryoprotectant demonstrated comparable efficacy to 10% dimethyl sulfoxide (DMSO).
The amino acid-based cryoprotectant achieved this at lower concentrations and with a convenient application method.

Conclusions:

A novel, non-toxic, amino acid-based cryoprotectant was successfully developed.
This formulation offers a promising DMSO-free alternative for cell cryopreservation in cell therapy.
The findings provide valuable insights for the advancement of green cryopreservation technologies.