Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: Active and latent viral phenotypes in HIV GKO-infected BBB pericytes. A , HIV GKO viral construct ( upper image ) and infection schematic diagram ( lower image ). B , representative images of the flow cytometric analysis illustrating the 3- and 5-days infections increasing in fluorescence. Uninfected cells (mKO2-/GFP-) are depicted in a blue gate , latently infected cells (mKO2+/GFP-) are depicted in an orange gate , and actively infected cells (mKO2/GFP+) are depicted in a green gate . C , D , and E , RNAscope analysis of HIV DNA, HIV RNA, and cell nuclei on 5-days HIV GKO infected pericytes sorted by fluorescence and cultured for 3 days. Image inlets show split colors for HIV DNA ( upper inlet ) and HIV RNA ( lower inlet ). Inlets focus on cell nuclei showing ( C ) no fluorescence in uninfected cells, ( D ) an increase of HIV DNA in latent cells, and ( E ) an increase of HIV RNA in actively infected cells, scale bars, 20 μm. F and H , RNAscope fluorescence intensity quantification graphs of HIV DNA and HIV RNA, respectively. G , HIV DNA copies per 1000 cells measured by qPCR using the CCR5 copy number for data normalization. I , HIVGag expression levels were measured by qPCR, and GAPDH was used as a housekeeping gene. Graphs indicate the mean ± SD from 3 to 5 independent experiments. ∗∗∗∗ p < 0.0001, ∗∗∗ p = 0.0002, ∗∗ p = 0.003, ∗ p < 0.0449.

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Infection, Construct, Fluorescence, Cell Culture, Expressing

Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: Transcriptomics signatures of latent and actively infected BBB pericytes. A , heat map analysis reveals the scaled expression of differentially expressed genes in BBB pericytes during active, latent, and uninfected stages. The list of differentially expressed genes depicted in this heat map is provided in Tables S1–S3 and all fold changes are compared to uninfected cells. B , Venn diagram analysis of actively and latently infected pericytes versus uninfected pericytes in green and orange respectively, and latently versus actively infected pericytes in light blue . C , D , and E , Volcano plots depict downregulated ( blue color ) and upregulated genes ( orange color ) in latent versus uninfected pericytes, actively infected versus uninfected pericytes, and latently versus actively infected BBB pericytes, respectively. Genes depicted as significant at a -log ( p -value) ≥ 2 and log (fold change) ≥ 0.5 are shown in color. The list of differentially expressed genes in these analyses is provided in Tables S1–S3 . All statistical analysis is prepared from pericyte libraries of three independent experiments.

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Infection, Expressing

Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: Canonical pathway and network analysis of DEGs within uninfected, latently, and actively infected BBB pericytes. A , C , and E , Ten most significant, differentially expressed, canonical pathways for actively infected versus uninfected, latent versus uninfected, and latent versus actively infected, respectively. A Z-score of ≥ 2 was defined as the threshold of significant activation, whilst Z-score ≤ 2 was defined as the threshold of significant inhibition. Upregulated pathways are depicted in orange and downregulated pathways are depicted in blue . B , D , and F , Overlapping canonical network analysis of actively infected versus uninfected, latent versus uninfected, and latent versus actively infected, respectively, depicting significant overlap of canonical pathways. Analysis is limited to the top 15 pathways by -log ( p -value) and lines are drawn for networks with a minimum of 5 overlapping interactors.

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Infection, Activation Assay, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: Estrogen receptor signaling pathway genes involved in host responses within uninfected, latently, and actively infected BBB pericyte populations. A , heat map revealing the scaled expression of DEGs in estrogen receptor signaling, PI3K signaling, EGF signaling, autophagy, and NFkB activation in BBB pericytes during latent and actively infected stages. B , Chord plot analysis identifying protein interactions of the estrogen receptor signaling network across pathways. C , AKT1-based network map predicting cell outcomes based on DEGs for latent vs actively infected pericyte populations. Map shows a predicted activation of proliferation and predicted inhibition of apoptosis in latent cells relative to actively infected cells. D , AKT1-based network analysis for actively infected versus uninfected populations. Map shows a predicted inhibition of proliferation and predicted activation of apoptosis in actively infected cells relative to uninfected cells, directly opposite to the outcome predicted in ( C ). Proteins predicted to downregulate are depicted in blue while proteins predicted to upregulate are depicted in orange .

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Infection, Expressing, Activation Assay, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: AKT affects latent HIV-1 reservoir size in BBB pericytes. BBB pericytes were infected with HIV GKO and exposed to AKT activator Yoda1, AKT inhibitor Honokiol, or DMSO (Vehicle). A–C , FACS analysis of mKO2 and GFP expression. Uninfected cells (mKO2-/GFP-) are depicted in a blue gate , latently infected cells are depicted in an orange gate , and actively infected cells are depicted in a green gate . A , FACS analysis of cells treated with Vehicle. B , FACS analysis of cells treated with Yoda1. C , FACS analysis of cells treated with Honokiol. D , quantification graph of the latent reservoir size across treatments. Graphs indicate the mean ± SD from 6 to 7 independent experiments. ∗∗∗ p = 0.0002, ∗∗ p = 0.003, ∗ p < 0.0449.

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Infection, Expressing

Journal: The Journal of Biological Chemistry

Article Title: AKT signaling modulates latent viral reservoir viability in HIV-1-infected blood–brain barrier pericytes

doi: 10.1016/j.jbc.2023.105526

Figure Lengend Snippet: AKT activity regulates the survival of latently infected cells through changes in apoptosis. A , representative FACS analysis of mKO2 and GFP expression in vehicle ( upper panel ), Yoda1 ( middle panel ), and Honokiol ( lower panel ) treated cells gated for early apoptosis (Annexin V+ DAPI –) and ( B ) late apoptosis (Annexin V+ DAPI +). Gated populations delineating uninfected, latently, and actively infected pericyte populations. C and D , cell number quantification of early and late apoptosis in latently infected cells. E and F , cell number quantification of early and late apoptosis in actively infected cells. Graphs indicate the mean ± SD from 6 to 9 independent experiments. ∗∗∗∗ p < 0.0001, ∗∗∗ p = 0.0002, ∗∗ p = 0.003, ∗ p < 0.0449.

Article Snippet: Primary human brain vascular pericytes (ScienCell, Cat# 1200) were maintained in a hormone depleted pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with 2% Fetal Bovine Serum, charcoal stripped, USDA-approved regions, One Shot format (Thermofisher Cat#A3382101), 0.2% pericyte growth supplement, 100 units/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Activity Assay, Infection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: Evaluation of blood–retinal barrier integrity by transendothelial electrical resistance (TEER) in monocultures of human retinal endothelial cells (HRECs) and in co-cultures of HRECs with adipose-derived mesenchymal stem cells (ASCs) or pericyte-like differentiated ASCs (P-ASCs). In the upper part of the figure, TEER measurement procedures are illustrated. HRECs were first seeded in the underside of transwell inserts. In some samples, glucose was added to mimic hyperglycemic (HG) conditions. After 48 h, ASCs or P-ASCs were also seeded on the topside. TEER values were initially measured after 24 h of co-culture (T0), and for the following 3 days. In some other samples, TEER measurements were carried out on corresponding cultures maintained in normal glucose (NG) conditions. TEER values were also measured in cultures of HRECs alone, in NG or HG conditions, at corresponding time points. The results obtained are reported in the graph in the lower part of the figure. For each sample, data are expressed as percentage changes with respect to the value measured at T0, assumed as a control. As was expected, no significant variations were observed in samples kept in NG conditions. Instead, different changes were found for the various HG samples: the most evident decreases were exhibited by HG HRECs; except for day 1, lower decreases were noticeable when HRECs were co-cultured with ASCs; minimal decreases were observed when P-ASCs were also present. In fact, in the last case, no significant variations were revealed at day 2 and 3, compared with NG conditions. Values are expressed as the mean ± standard error of the mean (SEM) of results from three independent experiments, with four parallel samples per group in each experiment. * p < 0.05 vs. T0 (starting point of measurements); ♦ p < 0.05 vs. NG respective controls (CTRL); § p < 0.05 vs. respective HG; # p < 0.05 vs. HRECs/ASCs-conditioned media. One-way ANOVA, followed by Tukey’s test. Created with BioRender.com.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Derivative Assay, Co-Culture Assay, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: Double-labeling immunofluorescence experiments in co-cultures of human retinal endothelial cells (HRECs) and human adipose-derived mesenchymal stem cells (ASCs) or pericyte-like ASCs (P-ASCs). Micrographs obtained after 1 day ( A – C ) and 4 days ( B – D ) of mono- or co-culture, in NG or in HG. Photos A1, B1, C1, and D1 refer to HREC monocultures, taken as a reference: only VE-cadherin ( A , B ) or ZO-1 ( B – D ) immunostainings are present. In both control NG-HRECs, at day 4 (B-1 and D-1), the immunofluorescence signals were higher than at day 1 (A-1 and C-1), revealing the greater time-dependent expressions of both junction proteins and confirming the characteristic intercellular contacts, typical of blood–retinal barrier cells. In NG, the fluorescent signals were significantly stronger when HRECs were in direct mixed cultures with ASCs (A2, B2, C2, and D2) and, even more, with P-ASCs (A3, B3, C3, and D3) for both proteins at the two time points. The fluorescence related to both VE-cadherin and ZO-1 reduced significantly in HG incubation (A4, B4, C4, and D4). However, strong recoveries were observed in HRECs conditioned by ASCs (VE-cadherin, A5 and B5; ZO-1, C5 and D5) and, more significantly, by P-ASCs (VE-cadherin, A6 and B6; ZO-1, C6 and D6). No α-SMA immunoreactivity is noticeable in A1-2, B1-2, C1-2, and D1-2, which refer to HRECs in monoculture. DAPI staining of cell nuclei is indicated by blue fluorescence. Overall, these results indicate that, even in HG concentrations, the presence of P-ASCs is able to preserve the integrity of junction proteins. Scale bar: 50 µm. Histograms on the right side of each panel refer to quantification of fluorescence intensity. Values are expressed as the mean ± SEM of results from three independent experiments, with four parallel samples per group in each experiment. * p < 0.05 vs. HRECs in NG; # p < 0.05 vs. HRECs in HG; § p < 0.05 HRECs/P-ASCs in HG vs. HRECs/ASCs in HG.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Labeling, Immunofluorescence, Derivative Assay, Co-Culture Assay, Fluorescence, Incubation, Staining

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: TNF-α, IL-1β, and VEGF mRNA levels in cultures of human retinal endothelial cells (HRECs) alone and co-cultured with human adipose-derived mesenchymal stem cells (ASCs) or pericyte-like ASCs (P-ASCs). TNF-α, IL-1β, and VEGF mRNA levels in HRECs were measured by qRT-PCR from samples that were cultured either in normal glucose (NG) or high glucose (HG) conditions. mRNA levels of each group were normalized to the housekeeping reference gene ribosomal 18S RNA. In the histograms, values are expressed as the fold change in those detected in HRECs cultured in NG conditions. Data indicate a modulation of inflammation genes in contrast to an HG-induced pro-inflammatory HREC phenotype. At the same time, decreases in pro-angiogenic factors were found. More evident effects were observed when HRECs were co-cultured with P-ASCs. Each value represents mean ± SEM obtained from three independent experiments. * p < 0.05 vs. HRECs in NG; # p < 0.05 vs. HRECs in HG; $ p < 0.05 HRECs/P-ASCs in HG vs. HRECs/ASCs in HG.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Cell Culture, Derivative Assay, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: TGF-β1 mRNA levels in cultures of human retinal endothelial cells (HRECs) alone and co-cultured with human adipose-derived mesenchymal stem cells (ASCs) or pericyte-like ASCs (P-ASCs). TGF-β1 mRNA levels were measured in HRECs by qRT-PCR from samples that were cultured either in normal glucose (NG) or high glucose (HG) conditions. mRNA levels of each group were normalized to the housekeeping reference gene ribosomal 18S RNA. In the histograms, values are expressed as the fold change in those detected in HRECs cultured in NG conditions. A significant reduction in HG-induced increased TGF-β1 mRNA levels can be found when HRECs were co-cultured with ASCs and, especially, P-ASCs. Each value represents mean ± SEM obtained from three independent experiments. * p < 0.05 vs. HRECs in NG; # p < 0.05 vs. HRECs in HG; $ p < 0.05 HRECs/P-ASCs in HG vs. HRECs/ASCs in HG.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Cell Culture, Derivative Assay, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: Evaluation of the ERK1/2-cPLA2-COX-2 axis in cultures of human retinal endothelial cells (HRECs) alone and co-cultured with human adipose-derived mesenchymal stem cells (ASCs) or pericyte-like ASCs (P-ASCs). Data were gathered by Western blot analysis from samples that were cultured either in normal glucose (NG) or high glucose (HG) conditions. Immunoblot analyses were performed in HREC populations using specific antibodies against p-ERK 1/2 and total ERK 1/2, activated (phosphorylated, p-cPLA2) and total cPLA2, and COX-2 proteins (panel A ). ß actin was used to verify the equal loading of 30 μg protein per lane. Image J software was used to carry out densitometric analysis of the immunoblots, indicating protein quantification of each band (in arbitrary densitometry units, a.d.u.). Quantitative analyses of the phosphorylation rate of ERK1/2 (i.e., ratio of p-ERK1/2/ERK1/2) and cPLA2 (i.e., ratio of p-cPLA2/cPLA2) are indicated in panels B and C , respectively. Quantitative analysis of COX-2 protein was normalized to ß actin (panel D ). No significant differences were noted among the different samples when kept in NG conditions. In HG conditions, HREC activation of the ERK1/2-cPLA2-COX-2 axis was attenuated by the presence of ASCs and, especially, P-ASCs. Each value represents mean ± SEM obtained from three independent experiments. * p < 0.05 vs. HRECs in NG; # p < 0.05 vs. HRECs in HG; § p < 0.05 HRECs/P-ASCs in HG vs. HRECs/ASCs in HG.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Cell Culture, Derivative Assay, Western Blot, Software, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: Evaluation of the PDGF-B/PDGFR-β axis in cultures of human retinal endothelial cells (HRECs) alone and co-cultured with human adipose-derived mesenchymal stem cells (ASCs) or pericyte-like ASCs (P-ASCs). By qRT-PCR, PDGF-B RNA levels were measured from HRECs (panel A ) and PDGFR-β mRNA levels were measured from ASCs or P-ASCs (panel B ). Cell samples that were cultured either in normal glucose (NG) or high glucose (HG) conditions were evaluated. mRNA levels of each group were normalized to the housekeeping reference gene ribosomal 18S RNA. Although the PDGF-B RNA levels were increased in cultures of HREC alone by glucose addition, much more evident increases were obtained when P-ASCs were also present, both in NG and HG conditions. On the other hand, when detecting PDGFR-β RNA levels in ASCs or P-ASCs, basal values observed in ASCs were greatly increased in P-ASCs, both in NG and HG conditions. Data indicate that enhanced pericyte recruitment would occur if P-ASCs are present. Each value represents mean ± SEM obtained from three independent experiments. Panel A : * p < 0.05 vs. HRECs in NG; # p < 0.05 vs. HRECs in HG; $ p < 0.05 HRECs/P-ASCs in HG vs. HRECs/ASCs in HG. Panel B : * p < 0.05 vs. the corresponding co-culture in NG.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Cell Culture, Derivative Assay, Quantitative RT-PCR, IF-P, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Protective Effects of Human Pericyte-like Adipose-Derived Mesenchymal Stem Cells on Human Retinal Endothelial Cells in an In Vitro Model of Diabetic Retinopathy: Evidence for Autologous Cell Therapy

doi: 10.3390/ijms24020913

Figure Lengend Snippet: Schematic diagram of blood–retinal barrier (BRB) integrity restored (preserved) by the treatment with pericyte-like differentiated adipose-derived mesenchymal stem cells (P-ASCs) in the context of diabetic retinopathy. ( A ) In normal glucose conditions, human retinal endothelial cells (HRECs) tightly adhere to each other by both adherens (VE-cadherin) and tight junctions (zonula occludens-1; ZO-1), providing protection to retinal structures. The presence of cell junctions is responsible for high transendothelial electrical resistance (TEER) values, which are indicative of BRB integrity. In a health context, the release of inflammatory cytokines (i.e., interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α)), pro-angiogenic factor, vascular endothelial growth factor-A (VEGF-A), and matrix metalloproteinase-9 (MMP-9) is limited to satisfying homeostatic needs only. HRECs produce and release the platelet-derived growth factor-β (PDGF-β), which is essential for the recruitment of pericytes to vessels. This molecule interacts with the platelet-derived growth factor receptor-β (PDGFR-β), located on the pericyte surface, triggering an intracellular signaling cascade that ensures their survival, proliferation, and migration. ( B ) In diabetic retinopathy, high glucose concentrations cause BRB breakdown, characterized by a series of events involving endothelial cells, pericytes, and the extracellular environment. In HRECs, high glucose levels induce the activation of the p-ERK1/2/p-cPLA2/COX-2 axis associated with the inflammatory state. Moreover, the connections between endothelial cells become weaker because of the downregulation of both adherens and tight junctions. This condition, revealed by lower TEER values, makes the BRB more accessible to external insults. Endothelial cells also overexpress VEGF-A, which leads to increased angiogenesis correlated to inflammation. Furthermore, the excess of glucose causes a reduction in PDGF-β release from HRECs, and a concomitant decrease in PDGFR-β expression on pericytes. As a result, pericyte population density decreases, leaving endothelial cells unprotected. At the extracellular level, hyperglycemia provokes an inflammatory state supported by the release of IL-1β and TNF-α, in addition to the increased release of MMP-9, responsible for the remodeling of the extracellular matrix. ( C ) Intraocular administration of P-ASC might result in beneficial effects, since these cells could act as substitutes for lost pericytes, thus limiting hyperglycemia-induced deleterious effects and possibly restoring BRB integrity.

Article Snippet: According to a previous study [ ], the pericyte like-differentiation of ASCs was achieved by their growth for 3 days in a culture medium specifically designed for pericytes (PM; Innoprot) containing 2% FBS and 5 mM glucose (normal glucose, NG).

Techniques: Derivative Assay, Migration, Activation Assay, Expressing