Optimizing CD34+ Cell Genome Editing for Efficiency and HSPC Maintenance

CRISPR-Cas9 gene editing of human hematopoietic stem and progenitor cells (HSPCs) holds great promise for disease treatment by cellular therapy and other methodologies. However, the variables that can affect treatment of relevant diseases in the real world are still being elucidated. A critical part of this involves implementing an optimized gene editing and cell culture methodology.

In this webinar, Dr. Marta Walasek and Danielle Nguyen Truong describe tips and tricks to lock down effective genome editing conditions for CD34+ cells, methods to evaluate genome editing efficiency, as well as optimal pre- and post-editing culture conditions to maintain HSPC function and long-term editing effects.

The speakers outline the differences between primary CD34+ cell sources and the advantages of using StemSpan™ serum-free cell culture media for the maintenance of CD34 expression and expansion of primitive HSPC subsets during genome editing.

Key considerations for designing CRISPR-Cas9 gene editing protocols are also reviewed by presenting a case study that demonstrates efficient genetic knockout of CD45 and β2 Microglobulin (B2M) in CD34+ HSPCs using the ArciTect™ CRISPR-Cas9 system.