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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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Pericyte-like differentiation of human adipose-derived mesenchymal stem cells:An in vitro study

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World Journal of Stem Cells 2020年第10期12卷 1152-1170页

作者： Giuliana Mannino Florinda Gennuso giovanni giurdanella Federica Conti Filippo Drago Salvatore Salomone Debora Lo Furno Claudio Bucolo Rosario Giuffrida Physiology Section Department of Biomedical and Biotechnological SciencesUniversity of CataniaCatania 95123Italy Pharmacology Section Department of Biomedical and Biotechnological SciencesSchool of MedicineUniversity of CataniaCatania 95123Italy Biochemistry Section Department of Biomedical and Biotechnological SciencesSchool of MedicineUniversity of CataniaCatania 95123Italy

BACKGROUND Adipose-derived mesenchymal stem cells(ASCs)are characterized by long-term self-renewal and a high proliferation *** adequate conditions,they may differentiate into cells belonging to mesodermal,endodermal or ectodermal *** support endothelial cells and play an important role in stabilizing the vessel wall at the microcirculation *** loss of pericytes,as occurs in diabetic retinopathy,results in a breakdown of the blood-retina barrier(BRB)and infiltration of inflammatory *** this context,the use of pericytelike differentiated ASCs may represent a valuable therapeutic strategy for restoring BRB *** To test in vitro strategies to obtain pericyte-like differentiation of human ASCs(hASCs).METHODS Different culture conditions were tested:hASCs cultured in a basal medium supplemented with transforming growth factorβ1;and hASCs cultured in a specific pericyte medium(PM-hASCs).In a further sample,pericyte growth supplement was omitted from the *** addition,cultures of human retinal pericytes(hRPCs)were used for ***-like differentiation of hASCs was tested by immunocytochemical staining and western blotting to evaluate the expression ofα-smooth muscle actin(α-SMA)and neural/glial antigen 2(NG2).Interactions between human retinal endothelial cells(hRECs)and different groups of hASCs were investigated in co-culture *** these cases,the expression of typical junctional proteins such as vascular endothelial-Cadherin,zonula occludens-1 and Occludin were assessed in *** an in vitro model of the BRB,values of trans-endothelial electrical resistance were measured when hRECs were co-cultured with various groups of pretreated *** values observed were compared with co-cultures of hRECs and hRPCs as well as with cultures of hRECs ***-dimensional co-cultures of hRECs and hRPCs or pericyte-like hASCs in Matrigel were designed to assess their reciprocal *** After 3-6 d of cult

关键词： Adipose-derived mesenchymal stem cells Pericyte-like differentiation Retinal endothelial cells Retinal pericytes Blood-retina barrier Junction proteins

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Potential therapeutic applications of mesenchymal stem cells for the treatment of eye diseases

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World Journal of Stem Cells 2021年第6期13卷 632-644页

作者： Giuliana Mannino Cristina Russo Anna Longo Carmelina Daniela Anfuso Gabriella Lupo Debora Lo Furno Rosario Giuffrida giovanni giurdanella Department of Biomedical and Biotechnological Sciences School of MedicineUniversity of CataniaCatania 95123Italy

Stem cell-based treatments have been extensively explored in the last few decades to develop therapeutic strategies aimed at providing effective alternatives for those human pathologies in which surgical or pharmacological therapies produce limited *** stem cells of different sources,mesenchymal stem cells(MSCs)offer several advantages,such as the absence of ethical concerns,easy harvesting,low immunogenicity and reduced tumorigenesis *** than a multipotent differentiation ability,MSCs can release extracellular vesicles conveying proteins,mRNA and *** to these properties,new therapeutic approaches have been designed for the treatment of various pathologies,including ocular *** this review,the use of different MSCs and different administration strategies are described for the treatment of diabetic retinopathy,glaucoma,and retinitis *** a large number of investigations,positive results have been obtained by in vitro experiments and by MSC administration in animal *** authors agree that beneficial effects are likely related to MSC paracrine *** on these considerations,many clinical trials have already been carried ***,although some adverse effects have been described,promising outcomes are *** can be assumed that in the near future,safer and more effective protocols will be developed for more numerous clinical applications to improve the quality of life of patients affected by eye diseases.

关键词： Mesenchymal stem cells Eye diseases Diabetic retinopathy Glaucoma Retinitis pigmentosa Regenerative medicine

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