Streamlined Methods for Processing and Cryopreservation of Cell Therapy Products Using Automated Systems

Ye Li; Hazel Y Stevens; Srikanth Sivaraman; Logan N Porter; Allison R Hoffman; Stuart L Gibb; Shivaram Selvam; Annie C Bowles-Welch

doi:10.21769/BioProtoc.4900

Streamlined Methods for Processing and Cryopreservation of Cell Therapy Products Using Automated Systems

Bio Protoc. 2023 Dec 20;13(24):e4900. doi: 10.21769/BioProtoc.4900.

Authors

Ye Li¹, Hazel Y Stevens¹, Srikanth Sivaraman¹, Logan N Porter¹, Allison R Hoffman², Stuart L Gibb², Shivaram Selvam¹, Annie C Bowles-Welch¹

Affiliations

¹ Marcus Center for Therapeutic Cell Characterization and Manufacturing, Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA.
² Terumo Blood Cell and Technologies, Lakewood, Colorado, USA.

Abstract

Streamlined procedures for processing and cryopreservation of cell therapies using good laboratory practices are integral to biomanufacturing process development and clinical applications. The protocol herein begins with the preparation of human cell types cultured as adherent (i.e., mesenchymal stromal cells, MSCs) or suspension cells (i.e., peripheral blood mononuclear cells, PBMCs) to comprehensively demonstrate procedures that are applicable to commonly used primary cell cultures. Cell processing steps consist of preparing high yields of cells for cryopreservation using instruments routinely used in cell manufacturing, including the Finia^® Fill and Finish System and a controlled-rate freezer. The final steps comprise the storage of cells at subzero temperatures in liquid nitrogen vapor phase followed by the analysis of cell phenotypes before and after processing and cryopreservation, along with cell quality metrics for validation. Additionally, the protocol includes important considerations for the implementation of quality control measures for equipment operation and cell handling, as well as Good Laboratory Practices for cell manufacturing, which are essential for the translational use of cell therapies. Key features • The protocol applies to small- or large-scale manufacturing of cell therapy products. • It includes streamlined procedures for processing and cryopreservation of cells cultured as adherent cells (MSCs) and suspension cells (PBMCs). • Provides temperature control and rapid partitioning of sample in cryopreservation solution to maintain high viability of a range of cell types throughout the procedures. • This protocol employs the Finia^® Fill and Finish System and a controlled-rate freezer. Graphical overview.

Keywords: Automated systems; Biomanufacturing; Cell therapies; Cryopreservation; Finia® Fill and Finish System; Mesenchymal stromal cells; Peripheral blood mononuclear cells; Process development.