title: Microporation is a valuable transfection method for efficient gene delivery into human umbilical cord blood-derived mesenchymal stem cells.
abstract: BACKGROUND
  Mesenchymal stem cells (MSCs) are an attractive source of adult stem cells for therapeutic application in clinical study. Genetic modification of MSCs with beneficial genes makes them more effective for therapeutic use. However, it is difficult to transduce genes into MSCs by common transfection methods, especially nonviral methods. In this study, we applied microporation technology as a novel electroporation technique to introduce enhanced green fluorescent protein (EGFP) and brain-derived neurotropfic factor (BDNF) plasmid DNA into human umbilical cord blood-derived MSCs (hUCB-MSCs) with significant efficiency, and investigated the stem cell potentiality of engineered MSCs through their phenotypes, proliferative capacity, ability to differentiate into multiple lineages, and migration ability towards malignant glioma cells.
  
  
  RESULTS
  Using microporation with EGFP as a reporter gene, hUCB-MSCs were transfected with higher efficiency (83%) and only minimal cell damage than when conventional liposome-based reagent (<20%) or established electroporation methods were used (30-40%). More importantly, microporation did not affect the immunophenotype of hUCB-MSCs, their proliferation activity, ability to differentiate into mesodermal and ectodermal lineages, or migration ability towards cancer cells. In addition, the BDNF gene could be successfully transfected into hUCB-MSCs, and BDNF expression remained fairly constant for the first 2 weeks in vitro and in vivo. Moreover, microporation of BDNF gene into hUCB-MSCs promoted their in vitro differentiation into neural cells.
  
  
  CONCLUSIONS
  Taken together, the present data demonstrates the value of microporation as an efficient means of transfection of MSCs without changing their multiple properties. Gene delivery by microporation may enhance the feasibility of transgenic stem cell therapy.
authors.affiliation: Department of Biomedical Science, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Jung Yeon
authors.initials: JY
authors.lastname: Lim
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Park
authors.forename: Chang Hyun
authors.initials: CH
authors.lastname: Jeong
authors.forename: Ji Hyeon
authors.initials: JH
authors.lastname: Oh
authors.forename: Seong Muk
authors.initials: SM
authors.lastname: Kim
authors.forename: Chung Hun
authors.initials: CH
authors.lastname: Ryu
authors.forename: Soon A
authors.initials: SA
authors.lastname: Park
authors.forename: Jae Geun
authors.initials: JG
authors.lastname: Ahn
authors.forename: Wonil
authors.initials: W
authors.lastname: Oh
authors.forename: Sin-Soo
authors.initials: SS
authors.lastname: Jeun
authors.forename: Jong Wook
authors.initials: JW
authors.lastname: Chang
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D005312:Fetal Blood / Q000166:cytology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D010957:Plasmids; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*
chemical_list: D019208:Brain-Derived Neurotrophic Factor; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
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