Low molecular weight polyethylenimine for efficient transfection of human hematopoietic and umbilical cord blood-derived CD34+ cells.
With the emerging role of hematopoietic stem cells as potential gene and cell therapy vehicles, there is an increasing need for safe and effective nonviral gene delivery systems. Here, we report that gene transfer and transfection efficiency in human hematopoietic and cord blood CD34+ cells can be enhanced by the use of low molecular weight polyethylenimine (PEI). PEIs of various molecular weights (800-750,000) were tested, and our results showed that the uptake of plasmid DNA by hematopoietic TF-1 cells depended on the molecular weights and the N/P ratios. Treatment with PEI 2K (m.w. 2000) at an N/P ratio of 80/1 was most effective, increasing the uptake of plasmid DNA in TF-1 cells by 23-fold relative to Lipofectamine 2000. PEI 2K-enhanced transfection was similarly observed in hematopoietic K562, murine Sca-1+, and human cord blood CD34+ cells. Notably, in human CD34+ cells, a model gene transferred with PEI 2K showed 21,043- and 513-fold higher mRNA expression levels relative to the same construct transfected without PEI or with PEI 25 K, respectively. Moreover, PEI 2K-treated TF-1 and human CD34+ cells retained good viability. Collectively, these results indicate that PEI 2K at the optimal N/P ratio might be used to safely enhance gene delivery and transfection of hematopoietic and human CD34+ stem cells.
Shin
Jee-Young
JY
CHA Research Institute, College of Medicine, Pochon CHA University, Pochon, Kyonggi-do 487-800, South Korea.
Suh
Dongchul
D
Kim
Jung Mogg
JM
Choi
Han-Gon
HG
Kim
Jung Ae
JA
Ko
Jung Jae
JJ
Lee
Yong Bok
YB
Kim
Jin-Seok
JS
Oh
Yu-Kyoung
YK
D018952:Antigens, CD34 / Q000097:blood*; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage; D005312:Fetal Blood / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D020014:K562 Cells; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection / Q000379:methods*
D018952:Antigens, CD34; D007376:Interleukin-2; D011094:Polyethyleneimine; D004247:DNA