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| 1 | Pericyte-like differentiation of human adipose-derived mesenchymal stem cells:An in vitro study显示文摘BACKGROUND Adipose-derived mesenchymal stem cells(ASCs)are characterized by long-term self-renewal and a high proliferation rate.Under adequate conditions,they may differentiate into cells belonging to mesodermal,endodermal or ectodermal lineages.Pericytes support endothelial cells and play an important role in stabilizing the vessel wall at the microcirculation level.The loss of pericytes,as occurs in diabetic retinopathy,results in a breakdown of the blood-retina barrier(BRB)and infiltration of inflammatory cells.In this context,the use of pericytelike differentiated ASCs may represent a valuable therapeutic strategy for restoring BRB damage.AIM 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 PM.In addition,cultures of human retinal pericytes(hRPCs)were used for comparison.Pericyte-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 experiments.In these cases,the expression of typical junctional proteins such as vascular endothelial-Cadherin,zonula occludens-1 and Occludin were assessed in hRECs.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 hASCs.The values observed were compared with co-cultures of hRECs and hRPCs as well as with cultures of hRECs alone.Three-dimensional co-cultures of hRECs and hRPCs or pericyte-like hASCs in Matrigel were designed to assess their reciprocal localization.RESULTS After 3-6 d of culture,α-SMA and NG2 immunocytochemistry showed that the closest pericyte-like phenotype was observed when hASCs were cultured in Pericyte Medium(PM-hASCs).In particular,α-SMA immunoreactivity,already visible at the basal level in pericytes and ASCs,was strongly increased only when transforming growth factor was added to the culture medium.NG2 expression,almost undetectable in most conditions,was substantially increased only in PMhASCs.Immunocytochemical results were confirmed by western blot analysis.The presence of pericyte growth supplement seems to increase NG2 expression rather thanα-SMA,in agreement with its role in maintaining pericytes in the proliferative state.In co-culture experiments,immunoreactivity of vascular endothelial-Cadherin,zonula occludens-1 and Occludin was considerably increased in hRECs when hRPCs or PM-hASCs were also present.Supporting results were found by trans-endothelial electrical resistance measurements,gathered at 3 and 6 d of co-culture.The highest resistance values were obtained when hRECs were co-cultured with hRPCs or PM-hASCs.The pericyte-like phenotype of PM-hASCs was also confirmed in three-dimensional co-cultures in Matrigel,where PM-hASCs and hRPCs similarly localized around the tubular formations made by hRECs.CONCLUSION PM-hASCs seem able to strengthen the intercellular junctions between hRECs,likely reinforcing the BRB;thus,hASC-based therapeutic approaches may be developed to restore the integrity of retinal microcirculation. | Giuliana Mannino Florinda Gennuso Giovanni Giurdanella Federica Conti Filippo Drago Salvatore Salomone Debora Lo Furno Claudio Bucolo Rosario Giuffrida | 2020 | World Journal of Stem Cells2020，12，10： | 3 |
| 2 | The effect of ganglioside on oxidation-induced permeability changes in lens and in epithelial cells of lens and retina显示文摘 | Bucolo C Lin LR Dang L | 1994 | Exp Eye Res1994，58，6： | 1 |
| 3 | Flurbiprofen- loaded acrylate polymer nanosuspensions for ophthalmic application 显示文摘 | PIGNATELLO R BUCOLO C SPEDALIERI G | 2002 | Biomaterials2002，23，15： | 1 |
| 4 | Does chaos work better than noise显示文摘 | Caponetto R Fortuna L | 2002 | Circuits and Systems Magazine IEEE2002，2，3： | 1 |
| 5 | Dopamine D (3) receptor as a new pharmacological target for the treatment of depression显示文摘 | LEGGIO GM SALOMONE S BUCOLO C | 2013 | Eur J Pharmacol2013，719，13： | 1 |
| 6 | Safety profile assessment of buflomedil: an overview of adverse reactions between 1975 and 2011 显示文摘 | Bucolo C Longo L Camillieri G | 2012 | Pharmacoepidemiol Drug Saf2012，21，11： | 1 |
| 7 | Eudragit RS100 nanosus- pensions for the ophthalmic controlled delivery of ibuprofen显示文摘 | Pignatello R Bucolo C Ferrara P | 2002 | Eur J Pharm Sci2002，16，12： | 1 |
| 8 | The PKCb/HuR/ VEGF pathway in diabetic retinopathy显示文摘 | Amadio M Bucolo C Leqqio GM | 2010 | Biochem Phar- macol2010，80，8： | 1 |
| 9 | Image processing for medical diagnosis using CNN 显示文摘 | ARENA P BASILE A BUCOLO M | 2003 | Nuclear Instruments & Methods in Physics Research Section A2003，，497： | 1 |
| 10 | A water-soluble carbon monoxide-releasing molecule ( CORM-3 ) lowers intraocular pressure in rabbits显示文摘 | Stagni E Privitera MG Bucolo C | 2009 | Br J Ophthalmol2009，93，2： | 1 |
| 11 | Does chaos work better than noise? 显示文摘 | BUCOLO M CAPONETrO R FORTUNA L | 2002 | Circuits and Systems Magazine IEEE2002，2，3： | 1 |
| 12 | Metal fume fever显示文摘 | Michele Malaguarnera Filippo Drago Giulia Malaguarnera Giovanni Li Volti Salvatore Salomone Filippo Caraci Fabio Galvano Marco Vacante Claudio Bucolo Mariano Malaguarnera | 2013 | The Lancet . 2013 (9885)2013，，9885： | 1 |
| 13 | Flurbiprofenloaded acrylate polymer nanosuspensions for ophthalmic application显示文摘 | Pignatello R Bucolo C Spedalieri G | 2002 | Biomaterials2002，23，15： | 1 |
| 14 | Eudragit RS100 nanosuspensions for the ophthalmic controlled delivery of ibuprofen显示文摘 | Pignatello R Bucolo C Ferrara P | 2002 | Eur Pharm Sci2002，16，： | 1 |
| 15 | The PKCβ/HuR/VEGF pathway in diabetic retinopathy显示文摘 | M. Amadio C. Bucolo G.M. Leggio F. Drago S. Govoni A. Pascale | 2010 | Biochemical Pharmacology2010，，8： | 1 |
| 16 | Eudragit RS100 nanosuspensions for the ophthalmic controlled delivery of ibuprofen显示文摘 | Pignatello R Bucolo C Ferrara P | 2002 | Eur J Pharm Sci2002，16，12： | 1 |
| 17 | Protective effects of a coumarin derivative in diabetic rats 显示文摘 | Bucolo C Ward KW Mazzon E | 2009 | Invest Oph- thalmol Vis Sci2009，50，8： | 1 |
| 18 | Preparation and characterization of eudragit retard nanosuspensions for the ocular delivery of cloricromene显示文摘 | Pignatello R Ricupero N Bucolo C | 2006 | AAPS PharmSciTech2006，7，1： | 1 |
| 19 | The PKCbeta/HuR/VEGF pathway in diabetic retinopathyp显示文摘 | Amadio M Bucolo C Leggio GM | 2010 | Biochem Pharmacol2010，80，8： | 1 |
| 20 | Eriodictyol prevents early retinal and plasma abnormalities in streptozotocin-induced diabetic rats显示文摘 | Bucolo C Leggio GM Drago F | 2012 | Biochem Pharm2012，84，4： | 1 |
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