Combination of Chemical and Neurotrophin Stimulation Modulates Neurotransmitter Receptor Expression and Activity in Transdifferentiating Human Adipose Stromal Cells
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Adipose stromal cells are promising tools for clinical applications in regeneration therapies, due to their ease of isolation from tissue and its high yield; however, their ability to transdifferentiate into neural phenotypes is still a matter of controversy. Here, we show that combined chemical and neurotrophin stimulation resulted in neuron-like morphology and regulated expression and activity of several genes involved in neurogenesis and neurotransmission as well as ion currents mediated by NMDA and GABA receptors. Among them, expression patterns of genes coding for kinin-B1 and B2, α7 nicotinic, M1, M3 and M4 muscarinic acetylcholine, glutamatergic (AMPA2 and mGlu2), purinergic P2Y1 and P2Y4 and GABAergic (GABA-A, β3-subunit) receptors and neuronal nitric oxide synthase were up-regulated compared to levels of undifferentiated cells. Simultaneously, expression levels of P2X1, P2X4, P2X7 and P2Y6 purinergic and M5 muscarinic acetylcholine receptors were down-regulated. Agonist-induced activity levels of the studied receptor classes also augmented during neuronal transdifferentiation. Transdifferentiated cells expressed high levels of neuronal β3-tubulin, NF-H, NeuN and MAP-2 proteins as well as increased ASCL1, MYT1 and POU3F2 gene expression known to drive neuronal fate determination. The presented work contributes to a better understanding of transdifferentiation induced by neurotrophins for a prospective broad spectrum of medical applications.
KeywordsMesenchymal stem cells Adipose stromal cells Human lipoaspirate Transdifferentiation Neuronal differentiation
This work was funded by a grant from the Brazilian funding agencies Fundação de Amparo à Pesquisa do Estado de São Paulo (São Paulo Research Foundation, FAPESP project No.2012/50880-4). C.L. is grateful for grant support from the São Paulo State Foundation FAPESP (project No. 2015/19128-2). H.U. acknowledges Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for fellowship support. A.A.N.’s and V.B.’s doctoral thesis research was supported by a fellowship from FAPESP (Proj. No. 2006/61286-5 and 2008/52334-1, respectively). I.C.N. acknowledges fellowship support from FAPESP (Proj. No. 2015/18730-0) for her postdoctoral research. R.L.P.’s doctoral thesis research was supported by a fellowship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Prof. José E. Krieger, Heart Institute (InCor), University of São Paulo Medical School, São Paulo, Brazil, is acknowledged for providing human patient adipose stromal cells. The physician Dr. Gustavo Gibin Duarte is acknowledged for liposuction procedures on patients.
Compliance with Ethical Standards
Disclosure of Potential Conflicts of Interest
Authors have no conflicts of interest to declare.
Research Involving Human Participants and/or Animals
Ethics’ Committee approval at the Heart Center of São Paulo was obtained (SDC Incor - 3005/07/080 - CAPPesq 0759/07).
Informed consent was obtained from all individual participants included in the study.
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