title: In vitro neuronal induction of adipose-derived stem cells and their fate after transplantation into injured mouse brain.
abstract: The effect of substances known as inducers of neuronal differentiation on cultured human and mouse adipose-derived mesenchymal stem cells (ASCs) and their fate after transplantation into the injured and ischemic mouse brains were studied. ASCs were isolated from the human and mouse adipose tissue. Inducers of neuronal differentiation included β-mercaptoethanol, glial cell line-derived neurotrophic factor (GNDF), brain-derived neurotrophic factor (BDNF), retinoic acid (RA), 5-azacytidine, as well as their combinations. Three days after the induction, the phenotype of the induced cells was analyzed using immunocytochemistry and real-time PCR assay for differential expression of specific genes. The induction efficiency was evaluated by the increased transcription of neuronal differentiation markers: nestin, β-III-tubulin (Tub-B), microtubule-associated protein 2 (MAP2), and neuron-specific enolase (ENO2). The expression of marker genes was tested by immunocytochemical analysis. ASC cultivation in the medium with RA or BDNF in combination with 5- azacytidine for a week increased the mRNA and protein levels of nestin, Tub-B, and ENO2. The transplantation of induced mouse ASCs into the mouse brain increased the lifespan of the cells relative to control uninduced cells and promoted their migration from the transplantation site to the recipient cerebral parenchyma. The transplantation of the induced cells into the mouse brain pre-exposed to endothelin- 1 promoted a more active cell migration into the surrounding ischemic brain tissue. Thus, ASC exposure to RA or BDNF in combination with 5-azacytidine elevated the transcription of the neuronal differentiation markers and improved the viability and integration of ASCs grafted into the mouse brain.
authors.affiliation: Institute of Gene Biology, Institute of Gene Biology, Russian Academy of Sciences, 34 Vavilov st. 119334, Russian Federation. lkorochkin@mail.ru
authors.forename: G
authors.initials: G
authors.lastname: Pavlova
authors.forename: T
authors.initials: T
authors.lastname: Lopatina
authors.forename: N
authors.initials: N
authors.lastname: Kalinina
authors.forename: E
authors.initials: E
authors.lastname: Rybalkina
authors.forename: Y
authors.initials: Y
authors.lastname: Parfyonova
authors.forename: V
authors.initials: V
authors.lastname: Tkachuk
authors.forename: A
authors.initials: A
authors.lastname: Revishchin
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000328:Adult; D000818:Animals; D001374:Azacitidine / Q000008:administration & dosage*; D001930:Brain Injuries / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000008:administration & dosage*; D002454:Cell Differentiation; D000089622:Doublecortin Domain Proteins; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000008:administration & dosage*; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008623:Mercaptoethanol / Q000008:administration & dosage*; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000378:metabolism; D020813:Receptor, trkB / Q000235:genetics; D014212:Tretinoin / Q000008:administration & dosage*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D000089622:Doublecortin Domain Proteins; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007381:Intermediate Filament Proteins; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D009479:Neuropeptides; C498585:TUBB3 protein, human; D014404:Tubulin; C462941:beta3 tubulin, mouse; D014212:Tretinoin; D008623:Mercaptoethanol; D020813:Receptor, trkB; D010751:Phosphopyruvate Hydratase; D001374:Azacitidine