Authors:

Mustapha AL Hassan

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ABSTRACT Cryopreservation is a critical technique for the storage and preservation of cells, including natural killer (NK) cells, particularly important for the use of these cell in cancer immunotherapy. Traditional cryoprotectants, particularly DMSO, have high toxicity and can introduce safety issues. This project aims to establish guidelines for the cryopreservation of NK cells and evaluate their viability and function under different cryopreservation conditions using novel cryoprotectants which can maintain safety and potency after thawing. The NK-92 cell line was used as the model for this study. Various cryoprotectants, including dimethyl sulfoxide (DMSO), ethylene glycol, glycerol, polyethylene glycol, and trehalose, were tested for their effectiveness in preserving NK cell viability. The cryopreservation media were prepared using different concentrations of cryoprotectants and fetal bovine serum (FBS). The NK-92 cells were frozen in liquid nitrogen for 24-72 hours and then thawed and cultured for more than five days. Viability assessments were performed using trypan blue staining. Additionally, non-DMSO formulations of cryoprotectants were prepared to compare their freezing performance against DMSO. The viability and functionality of the thawed NK cells were evaluated and compared to fresh, non-frozen controls. The results of this study will provide valuable insights into the optimal cryopreservation conditions for maintaining the viability and function of NK cells, which will contribute to their successful application in research and clinical settings.

Source:

Purdue University / 2023

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Co-authors:

Mustapha AL Hassan