SCGT event 2024 - CN (2024)

Non-viral genome editing in Hematopoietic Stem Cells (HSCs) for cell-therapy applications - from discovery to manufacturing

Abstract

Optimizing electroporation conditions with the right parameters is crucial to ensure efficient and successful transfection of cells, maximizing the desired outcomes of gene editing or other applications.

The Invitrogen™ Neon™ NxT Electroporation System scalability to the large-scale Gibco CTS™ Xenon™ electroporation system offers researchers a versatile platform for small-scale optimization, enabling a smooth transition from initial experiments to large-scale applications

The cell processing and delivery instruments together with gene editing reagents and optimized gene delivery protocols developed through this work offers a robust solution for efficient and precise engineering of hematopoietic stem cells (HSCs) in various research and therapeutic applications.

Scalable solutions for expansion of non-virally generated CAR T cells

Abstract

Biomanufacturers rely on bioreactors with closed designs to grow genetically modified immune cells for cell therapy development, as they offer reduced risk of contamination. Generally, they use static bioreactors or those that agitate cells through rocking or stirring.

In a previous application note, it was demonstrated that expansion of unedited T cells using automated stirred-tank (STR) bioreactors was improved compared to static bioreactors.

This study aims to investigate the impact of expanding genetically edited T cells in a stirred tank bioreactor compared to a static bioreactor.

Feasibility of Wash and Formulation of Mesenchymal Stem Cells in Low-volume for Intra-tumoral Delivery Using Closed Counterflow Centrifugation

Abstract

Purpose: Pre-clinical development of a GMP-compliant, closed process, to wash and formulate modified human Mesenchymal Stem Cells (hMSCs) in low harvest volumes for intratumoral administration, with specific interest for recurrent Glioblastoma (GBM) treatment.

Methods: Human MSCs are expanded and engineered with a GFP expressing construct using NUS’s proprietary non-viral gene delivery method in 5-chamber CellStack, generating 0.5 billion engineered hMSCs. Prior to cryopreservation, the transfection efficiency and cell viability were determined by Flow Cytometry and NC-3000 nucelocounter, respectively. To remove DMSO from the cell product, CTS™ Rotea™ counterflow centrifuge system was evaluated. A 10-minute cell-wash and harvest protocol was applied. Excipient-grade Hartmann’s solution was used as a wash and the hypothermosol was used as formulation buffer. To assess cell quality and functionality, hMSCs were stored at 4degC for up to 4 hours.

Results: Cell harvest volumes ranged from 2.9mL – 3.5mL is possible. At various number of cell input (80-220 x 106), viable cell recovery was >60%. At cell viability >80%, the % of transfected cells, phenotypic characteristic and the anti-cancer potency of the engineered MSCs remain unchanged.