Aggregate formation in cryopreserved leukocyte suspensions

Area of Science:

• Biomedical Science
• Cell Biology
• Cryobiology

Background:

• Cryopreservation of leukocytes often results in significant granulocyte injury.
• This injury leads to cell aggregation and reduced cell recovery after preservation.
• Aggregate formation is exacerbated by the presence of serum in cryoprotective media.

Purpose of the Study:

• To investigate the factors contributing to aggregate formation during leukocyte cryopreservation.
• To evaluate the efficacy of sugar-containing washing solutions in mitigating cell aggregation and loss.

Main Methods:

• Leukocyte cryopreservation was performed with varying cryoprotective media.
• The effect of serum presence on aggregate formation was analyzed.
• Washing solutions with different concentrations of sugars (sucrose, lactose, dextrose) were tested.

Main Results:

• Aggregate formation primarily occurred in the presence of serum and involved mainly leukocytes.
• The addition of sugars to the washing solution significantly reduced aggregate formation.
• The reduction in aggregates was directly proportional to the final sugar concentration.

Conclusions:

• Granulocyte injury during cryopreservation is a key factor in cell loss and aggregation.
• Sugars in washing solutions are effective in preventing leukocyte aggregates, improving cell preservation.
• Serum potentiates aggregation, but fibrinogen presence is not the cause.