A new method for transduction of mesenchymal stem cells using mechanical agitation.
Applications of bone marrow-derived mesenchymal stem cells in gene therapy have been hampered by the low efficiency of gene transfer to these cells. In current transduction protocols, retrovirus particles with foreign genes make only limited contact with their target cells by passive diffusion and have short life spans, thereby limiting the chances of viral infection. We theorized that mechanically agitating the virus-containing cell suspensions would increase the movement of viruses and target cells, resulting in increase of contact between them. Application of our mechanical agitation for transduction process has increased the absorption of retrovirus particles more than five times compared to the previous static method without changing cell growth rate and viability. The addition of a mechanical agitation step increased transduction efficiency to 42%, higher than that of any other previously-known static transduction protocol.
Park
Jin-O
JO
Department of Microbiology and Immunology and Institute for Medical Science and Research Center for Industrial Development of BioFood Materials, Chonbuk National University Medical School, Chonbuk 561-712, Korea.
Park
Sung-Hoon
SH
Hong
Seong-Tshool
ST
D000042:Absorption / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000473:pathology / Q000502:physiology*; D002478:Cells, Cultured; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D055595:Mechanical Phenomena; D059630:Mesenchymal Stem Cells / Q000473:pathology / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic / Q000295:instrumentation / Q000379:methods*; D014771:Virion / Q000235:genetics