Low-oxygen-affinity red cells produced in a large-volume, continuous-flow electroporation system

Area of Science:

• Biotechnology
• Hematology
• Biomedical Engineering

Background:

• Human red blood cells modified with inositol hexaphosphate (IHP) exhibit reduced oxygen affinity.
• These IHP-modified cells maintain oxygen-carrying capacity while improving tissue oxygen delivery.
• Efficient production of IHP-containing red cells is crucial for potential clinical applications.

Purpose of the Study:

• To devise an efficient system for large-scale production of inositol hexaphosphate (IHP)-containing red blood cells.
• To optimize conditions for IHP encapsulation and red blood cell survival.
• To develop a procedure suitable for transfusion-ready cell preparation.

Main Methods:

• Static electroporation techniques were explored to determine optimal IHP incorporation and cell survival parameters.
• A flow electroporation system was developed and optimized based on static electroporation findings.
• Specific electrical pulse parameters (field strength, pulse length) and cooling were applied to red cells in IHP solution.

Main Results:

• Optimal IHP encapsulation and cell integrity were achieved using three exponential pulses (2.98 kV/cm, 2.0 msec) in a cooled flow electroporation chamber.
• Post-storage (24 hours, 37°C), cells demonstrated over 85% survival with hematologic indices comparable to controls.
• The p50 value of the modified cells doubled to 50.4 torr, indicating significantly lower oxygen affinity.
• Processing time for one unit of blood was reduced to 90 minutes.

Conclusions:

• The described system efficiently produces low-oxygen-affinity red blood cells suitable for clinical transfusion.
• This method offers a viable approach for generating large volumes of modified red cells for therapeutic use.
• The optimized electroporation process balances IHP incorporation, cell viability, and processing efficiency.