An efficient gene transfer system for hematopoietic cell line using transient and stable vectors

The hematopoietic system represents an interesting model for gene transfer protocols. Here, we have evaluated the efficiency of a gene transfer system using the polycationic compound SuperFect (Qiagen) and the K562 hematopoie tic cell line. Transient and stable vectors carrying the enhanced green flu orescent protein (EGFP) reporter gene were employed. The stable vector was constructed based on Epstein-Barr virus sequences such as EBV oriP (origin of replication) and EBNA (EBV nuclear antigen)-1, both for DNA replication. The transfection efficiency of the viable cells was estimated by flow cyto metry at approximately 98% for transient and stable vectors. Transiently tr ansfected cells presented optimal EGFP expression until day 2 when fluoresc ence started to decrease. In contrast, stable transfectants continuously ex pressed the marker gene product for 10 weeks in the presence of G418. Our r esults represent an efficient gene transfer method for K562 hematopoietic c ells and may be used as an alternative approach for further gene transfer s tudies involving hematopoietic cells. (C) 2001 Elsevier Science B.V. All ri ghts reserved.