This presentation was given during the ISCT – International Society for Cell & Gene Therapy May, 2022.

Abstract:

In the past, engineered keratinocyte-based cell therapies have been limited by the lack of efficient transfection methods for adult keratinocytes. Here we aimed to develop a GMP-compliant, scalable cell engineering process using the MaxCyte^® cell electroporation platform to transfect neonatal and adult primary keratinocytes from four distinct anatomical locations. Delivery of multiple CRISPR RNPs in a single electroporation achieved highly efficient, multiplexed gene editing in a simple, adaptable process.

These improvements in transfection efficiency and cell viability reduced keratinocyte engineering times by up to 4 weeks, with a significantly higher success rate than a standard chemical transfection method.

Lastly, we demonstrate the scalability of the MaxCyte electroporation process, enabling the engineering of millions of primary keratinocytes without any loss of efficacy.