New technology cuts down on cell culture time by half and uses more targeted cell sorting and purification methods, which may help to address the issue of insufficient blood supply faced by healthcare systems around the world.
Researchers from the Singapore-MIT Alliance for Research and Technology (SMART) MIT’s research enterprise in Singapore, have discovered a new way to manufacture human red blood cells (RBCs) that cuts the culture time in half. Furthermore, they have developed novel purification and sorting techniques that are faster, more precise, and cheaper than existing methods.
Blood transfusions are an incredibly important resource in every healthcare system, saving millions of lives every year. However, they are often in limited supply in low- or middle- income countries, with an estimated 61 percent of countries not having enough supply to meet demand. The ability to manufacture RBCs on demand, particularly the universal donor blood (O+) would significantly benefit patients in need of transfusion by circumventing the need for large volume blood draws and difficult cell isolation processes.
In addition, being able to manufacture RBCs easily and quickly would also have a significant and positive impact on blood banks worldwide by reducing their dependence on donor blood, which has a higher risk of infection. Supplies of RBCs are also critical for research of diseases such as malaria and can also be used to develop new and improved cell therapies.
However, the existing methods for manufacturing and purifying RBCs are time-consuming and costly. They also produce undesirable by-products and are not optimal for large-scale therapeutic applications. A team from SMART’s Antimicrobial Research (AMR) and Critical Analytics for Manufacturing Personalised Medicine (CAMP), co-led by Principal Investigators Professor Jongyoon Han (MIT) and Professor Peter Preiser (NTU) were able to make huge technical advancements in the field of RBC manufacturing, explained in their paper, “Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture,” published recently in Lab on a Chip.
“Traditional methods for producing human RBCs usually require 23 days for the cells to grow, expand exponentially and finally mature into RBCs,” says Dr Kerwin Kwek, lead author of the paper and Senior Postdoctoral Associate at SMART CAMP. “Our optimised protocol stores the cultured cells in liquid nitrogen on what would normally be Day 12 in the typical process, and upon demand thaws the cells and produces the RBCs within 11 days.”
The team also developed new sorting and purification techniques by modifying existing Dean Flow Fractionation (DFF) and Deterministic Lateral Displacement (DLD) methods, leveraging the RBC’s size and deformability for purification instead of spherical size. As most human cells are deformable, this technique can have wide biological and clinical applications such as cancer cell and immune cell sorting and diagnostics.
Compared with conventional cell purification by fluorescence-activated cell sorting (FACS), SMART’s enhanced DFF and DLD methods were found to offer comparable purity while processing at least twice as many cells per second at less than a third of the cost. In scale-up manufacturing processes, DFF is more optimal for its high volumetric throughput, whereas in cases where cell purity is pivotal, DLD's high precision feature is most advantageous. The purified RBCs were found to retain their cellular functionality upon testing.
“Our novel sorting and purification methods result in significantly faster cell processing time and can be easily integrated into current cell manufacturing processes. The process also does not require a trained technician to perform sample handling procedures and is scalable for industrial production,” Dr Kwek added.
The results of their research would give scientists faster access to final cell products that are fully functional with high purity at a reduced cost of production.