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ARPE-19 conditioned medium promotes neural differentiation of adipose-derived mesenchymal stem cells

World Journal of Stem Cells 2021年第11期13卷 1783-1796页

作者： Giuliana Mannino Martina Cristaldi Giovanni Giurdanella Rosario Emanuele Perrotta Debora Lo Furno Rosario Giuffrida Dario Rusciano Department of Biomedical and Biotechnological Sciences University of CataniaCatania 95123CTItaly Research Center SOOFT-Italia S.p.A.Catania 95123CTItaly Department of General Surgery and Medical-Surgery Specialties University of CataniaCatania 95100CTItaly

BACKGROUND Adipose-derived stem cells(ASCs)have been increasingly explored for cell-based medicine because of their numerous advantages in terms of easy availability,high proliferation rate,multipotent differentiation ability and low *** this respect,they have been widely investigated in the last two decades to develop therapeutic strategies for a variety of human pathologies including eye *** ocular diseases involving the retina,various cell types may be affected,such as Müller cells,astrocytes,photoreceptors and retinal pigment epithelium(RPE),which plays a fundamental role in the homeostasis of retinal tissue,by secreting a variety of growth factors that support retinal *** To test ASC neural differentiation using conditioned medium(CM)from an RPE cell line(ARPE-19).METHODS ASCs were isolated from adipose tissue,harvested from the subcutaneous region of healthy donors undergoing liposuction *** ASC culture conditions were investigated:ASCs cultured in basal Dulbecco's Modified Eagle Medium(DMEM);ASCs cultured in serum-free DMEM;ASCs cultured in serumfree DMEM/F12;and ASCs cultured in a CM from ARPE-19,a spontaneously arising cell line with a normal karyotype derived from a human *** proliferation rate and viability were assessed by crystal violet and MTT assays at 1,4and 8 d of *** the same time points,ASC neural differentiation was evaluated by immunocytochemistry and western blot analysis for typical neuronal and glial markers:Nestin,neuronal specific enolase(NSE),protein gene product(PGP)9.5,and glial fibrillary acidic protein(GFAP).RESULTS Depending on the culture medium,ASC proliferation rate and viability showed some significant ***,less dense populations were observed in serum-free cultures,except for ASCs cultured in ARPE-19 serum-free ***,a different cell morphology was seen in these cultures after 8 d of treatment,with more elongated cells,often showing cytoplasmic ramifications.

关键词： Adipose-derived mesenchymal stem cells Retinal pigment epithelium neural markers neural differentiation Retina damage Cell-based medicine

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