High-level transgene expression in primary human T lymphocytes and adult bone marrow CD34+ cells via electroporation-mediated gene delivery

The design of effective gene delivery systems for gene transfer in primary human blood cells is important both for fundamental hematopoiesis research and for cancer gene therapy strategies.

Here, we evaluated electroporation as a nonviral means for transfection of activated human T lymphocytes and adult bone marrow (BM) CD34+ cells. We describe optimal culture and

electroporation parameters for efficient gene delivery in prestimulated T lymphocytes (16.3 ± 1.3%), as well as 2-day cultured adult BM CD34+ cells (29.6 ± 4.6%). PHA-stimulated T cells were

most receptive for transfection after 48h of in vitro culture, while T cells stimulated by CD3 cross-linking and interleukin (IL)-2 achieved maximum transfection levels after 72 h of

prestimulation. Kinetic analysis of EGFP expression revealed that activated T lymphocytes maintained transgene expression at high levels for a prolonged period. In addition, fresh

unstimulated BM CD34+ cells were consistently transfected (5.2 ± 0.4%) with minimal cytotoxicity (