Presentation: Scalable Electroporation of Adult Keratinocytes with Multiplexed CRISPR

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


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.



Andrew Mancini

Field Application Scientist

Andrew Mancini is a Field Application Scientist at MaxCyte specializing in Cell and Gene Therapy. As a field-based scientist, Andrew works closely with MaxCyte’s partners around the world to develop novel cell engineering workflows for the development of cellular therapeutics. Prior to his time at MaxCyte, Andrew completed his PhD in Biomedical Sciences at the University of California, San Francisco and completed his postdoctoral training at Genentech in Molecular Oncology. He has broad range of experience and expertise that encompasses non-viral and viral engineering of primary immune cells, stem cells, and cell lines at both research and manufacturing scales.


Céline Mahieu

Graduate Student, Aaron Tward lab UCSF

Céline Mahieu is currently a graduate student at the University of California, San Francisco (UCSF) pursuing her doctorate in Molecular Oncology. Prior to starting her doctoral work, she received her Bachelors and Masters degree in Biomedical Sciences from Utrecht University in the Netherlands. Under the co-supervision of Dr. Aaron Tward (UCSF) and Dr. Rene Bernards (Netherlands Cancer Institute), Celine’s doctoral research is focused on elucidating molecular mechanisms of squamous cell carcinoma pathogenesis. As part of this research, Céline has developed a novel method for the genome engineering of primary keratinocytes, piquing her interest in the development of novel epithelial based cell therapies.