Highly efficient transfection of human marrow stromal cells by nucleofection.
Human marrow stromal cells (hMSCs) are an attractive source for autologous cell and gene therapies. In this study, we developed a highly efficient transfection method for hMSCs. Although they tend to show efficient gene delivery, nonviral vectors offer several advantages over viral vectors for gene therapies. They are inexpensive to produce and suitable to adopt particularly with respect to little or no specific immune responses; they are simple to use; they entail easier large-scale production; and they have a high degree quality control. hMSCs and rat marrow stromal cells were transfected with the plasmid pEGFP-N1 that encoded a green fluorescent protein component by using two nonviral methods: nucleofection and electroporation. Nucleofection provided a much better rate of transfer than electroporation particularly in hMSCs.
Nakashima
S
S
Department of Orthopaedic Surgery, Nagoya University School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. shojiro@med.nagoya-u.ac.jp
Matsuyama
Y
Y
Nitta
A
A
Sakai
Y
Y
Ishiguro
N
N
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D003584:Cytological Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins