title: Improved lentiviral transduction of human mesenchymal stem cells for therapeutic intervention in pancreatic cancer.
abstract: Genetic modification of human bone marrow mesenchymal stem cells (MSC) is highly valuable for their exploitation in basic science and therapeutic applications, for example in cancer. We present here a new, fast and easy-to-use method to enrich a functional population of lentiviral (LV)-transduced MSC expressing enhanced green fluorescent protein (eGFP). We replaced the eGFP gene by a fusion gene of puromycin acetyltransferase and eGFP. Upon LV gene transfer and puromycin selection, we quickly obtained a pure transduced MSC population, in which growth, differentiation capacity and migration preferences were not compromised. Furthermore, we are the first to report the migration velocity of MSC among which 30% were moving and velocity of about 15 mum h(-1) was not altered by LV transduction. Manipulated MSC underwent senescence one passage earlier than non-transduced cells, suggesting the use for therapeutic intervention in early passage numbers. Upon tail vein application in nude mice, the majority of LV-transduced MSC could be detected in human orthotopic pancreatic tumor xenografts and to a minor extent in mouse liver, kidney and lung. Together, LV transduction of genes to MSC followed by puromycin selection is a powerful tool for basic research and improves the therapeutic prospects of MSC as vehicles in gene therapy.
authors.affiliation: Molecular OncoSurgery, University of Heidelberg and German Cancer Research Center, Heidelberg, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Kallifatidis
authors.forename: B M
authors.initials: BM
authors.lastname: Beckermann
authors.forename: A
authors.initials: A
authors.lastname: Groth
authors.forename: M
authors.initials: M
authors.lastname: Schubert
authors.forename: A
authors.initials: A
authors.lastname: Apel
authors.forename: A
authors.initials: A
authors.lastname: Khamidjanov
authors.forename: E
authors.initials: E
authors.lastname: Ryschich
authors.forename: T
authors.initials: T
authors.lastname: Wenger
authors.forename: W
authors.initials: W
authors.lastname: Wagner
authors.forename: A
authors.initials: A
authors.lastname: Diehlmann
authors.forename: R
authors.initials: R
authors.lastname: Saffrich
authors.forename: U
authors.initials: U
authors.lastname: Krause
authors.forename: V
authors.initials: V
authors.lastname: Eckstein
authors.forename: J
authors.initials: J
authors.lastname: Mattern
authors.forename: M
authors.initials: M
authors.lastname: Chai
authors.forename: G
authors.initials: G
authors.lastname: Schütz
authors.forename: A D
authors.initials: AD
authors.lastname: Ho
authors.forename: M M
authors.initials: MM
authors.lastname: Gebhard
authors.forename: M W
authors.initials: MW
authors.lastname: Büchler
authors.forename: H
authors.initials: H
authors.lastname: Friess
authors.forename: P
authors.initials: P
authors.lastname: Büchler
authors.forename: I
authors.initials: I
authors.lastname: Herr
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D010190:Pancreatic Neoplasms / Q000473:pathology / Q000628:therapy*; D014161:Transduction, Genetic
chemical_list: D049452:Green Fluorescent Proteins