Exogenous Nurr1 gene expression in electrically-stimulated human MSCs and the induction of neurogenesis.
In this study, synergistic effects of electrical stimulation and exogenous Nurr1 gene expression were examined to induce the differentiation of human mesenchymal stem cells (hMSCs) into nerve cells in in vitro culture system. A two-step procedure was designed to evaluate the effects of electrical stimulus and exogenous gene delivery for inducing neurogenesis. First, an electrical stimulation device was designed using gold nanoparticles adsorbed to the surface of a cover glass. Gold nanoparticles, as an electrical conductor for stem cells, are well-defined particles adsorbed to a polyethyleneimine (PEI)-coated cover glass. The nanoparticle morphology was examined by scanning electron microscope (SEM). Second, a plasmid carrying Nurr1 cDNA was complexed with biodegradable poly-(DL)-lactic-co-glycolic acid (PLGA) nanoparticles to support neurogenesis. To evaluate the neuronal differentiation of stem cells mediated by the treatment with either electrical stimulation and exogenous Nurr1 gene delivery, or both, the expression of neuron-specific genes and proteins was examined by RT-PCR and Western blotting. Cells transfected with exogenous Nurr1 genes plus electrical stimulation (250 mV for 1000 s) showed the greatest level of neurite outgrowth with a mean neurite length of 150 μm. Neurite length in cells treated with only one stimulus was not significant, approximately 10-20 μm. These results indicate that electrical stimulation and exogenous Nurr1 gene expression together may be adequate to induce nerve regeneration using stem cells.
Department of Biomedical Science, College of Life Science, CHA University, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea.
Ji Sun
JS
Park
Han Na
HN
Yang
Dae Gyun
DG
Woo
Su Yeon
SY
Jeon
Hyun-Jin
HJ
Do
Sun-Hyung
SH
Huh
Nam-Hyung
NH
Kim
Jae-Hwan
JH
Kim
Keun-Hong
KH
Park
D001672:Biocompatible Materials / Q000737:chemistry; D002470:Cell Survival; D018076:DNA, Complementary / Q000378:metabolism; D005786:Gene Expression Regulation; D006046:Gold / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D019344:Lactic Acid / Q000737:chemistry; D008027:Light; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D053768:Metal Nanoparticles / Q000737:chemistry; D008855:Microscopy, Electron, Scanning / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000096:biosynthesis* / Q000235:genetics*; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012542:Scattering, Radiation; D014162:Transfection
D001672:Biocompatible Materials; D018076:DNA, Complementary; C539882:NR4A2 protein, human; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D006046:Gold