MICROFLUIDIC SEPARATION TECHNOLOGY IMPROVES PURITY AND YIELD OF PBMCS FOR CELL MANUFACTURING APPLICATIONS

• Published in: Cytotherapy (Oxford, England) Vol. 26; no. 6; p. S26
• Main Authors: Song, H.W., Underwood, S., Traynor, R., Prochazkova, M., Cai, Y., Shah, N.N., Jin, P., Somerville, R., Stroncek, D., Highfill, S.L.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-06-2024
• Subjects: Cell isolation; MNC apheresis; PBMC; PBMC; Cell isolation; MNC apheresis
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1016/j.jcyt.2024.03.043

Mononuclear cell (MNC) apheresis products serve as the starting material of many cell therapies, and they can be further processed to yield purified peripheral blood mononuclear cells (PBMCs). This PBMC purification step eliminates excess red blood cells (RBCs) and platelets, since these can inhibit the culture and/or expansion of specific cell types of interest such as T cells, NK cells, or monocytes. The use of Ficoll density gradient separation efficiently separates PBMCs from RBCs, and this process has been adapted to the automated Sepax C-Pro device (Cytiva) for clinical manufacturing. In contrast to the Sepax, the Curate Cell Processing System uses microfluidic technology to separate PBMCs without the use of Ficoll.
We performed a series of side-by-side PBMC purifications using the Sepax and Curate systems with 6 healthy donor MNC apheresis products. The Curate microfluidic system yielded ≥75% recovery of total nucleated cells (average = 78.7 ± 2.7%), which was a 48% increase compared to Sepax, with a 77% increase in recovery of CD3+ T cells in particular. This improvement in recovery was accompanied by a significant decrease in both RBC and platelet contamination, with a 99.7 ± 0.1% reduction of platelets in the Curate compared to 90.9 ± 0.6% in the Sepax. Characterization of T cells following isolation showed an equivalent T cell differentiation state of naïve vs. effector cells between the two platforms, and current experiments are underway to evaluate performance of isolated PBMCs in a CAR-T manufacturing process. Given the improved recovery and purity of PBMCs isolated from the Curate microfluidic system, this technology demonstrates significant utility particularly for applications where large cell numbers and efficient cell recovery are desired.