Journal: Cell Death & Disease

Article Title: Direct conversion of human umbilical cord mesenchymal stem cells into retinal pigment epithelial cells for treatment of retinal degeneration

doi: 10.1038/s41419-022-05199-5

Figure Lengend Snippet: Pigment generation, RPE-specific and EMT-associated markers in iPSC-RPE cells, iRPE cells, and ARPE19 cells were detected by A pigment generation and immunostaining, B Western blotting and C quantitative analysis ( n = 3). D The Pearson correlation coefficient matrix of all samples based on RNA-seq datasets. E Electron micrographs of iPSC-RPE cells, iPRE cells, and ARPE19 cells. F Total bound and phagocyted POSs (pointed by arrows) in iPSC-RPE cells, iRPE cells, and ARPE19 cells. G Quantification of phagocytosis, as determined by the number of bound and phagocyted POS per field ( n = 8). H Tight junction were demonstrated by ZO-1 immunostaining. I TER analysis ( n = 4). J HRP permeability assay ( n = 4). K Expression levels of PEDF and VEGF from upper and lower chambers were determined by ELISA ( n = 4). Scale bar = 50 μm. Results are expressed as mean ± SD. P value measured by one-way ANOVA and post hoc Bonferroni’s test.

Article Snippet: PEDF and VEGF were quantified by PEDF ELISA kit (Elabscience, Wuhan, China) and VEGF ELISA kit (Proteintech).

Techniques: Immunostaining, Western Blot, RNA Sequencing Assay, Permeability, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Direct conversion of human umbilical cord mesenchymal stem cells into retinal pigment epithelial cells for treatment of retinal degeneration

doi: 10.1038/s41419-022-05199-5

Figure Lengend Snippet: A Experiment design for iRPE transplantation in SI-induced rat AMD model. B ERG waveforms recorded at different time points (the calibration indicates 200 μV vertically and 25 ms horizontally). C Quantitative analysis of ERG b-wave amplitude ( n = 10, P value measured by one-way ANOVA and post hoc Bonferroni’s test). D Representative micrographs of retinal samples at week 6 post-transplantation. The injection sites were pointed by arrows, and ONL was between yellow dashed lines. E Quantitative analysis of ONL thickness (μm) ( n = 6, P value measured by one-way ANOVA and post hoc Bonferroni’s test). F Representative micrographs of retinal cryosections stained with TUNEL. G Statistical analysis of the percentage of the apoptotic cells in ONL ( n = 6, P value measured by one-way ANOVA and post hoc Bonferroni’s test). H Immunostaining of hUCMSCs and iRPE cells after transplantation in vivo for 4 weeks. I Quantitative analysis of percent of immunostaining+ cells of grafted cells ( n = 7, P value measured by Student’s unpaired t test). J The level of PEDF secreted by hUCMSCs and iRPE cells were determined by ELISA ( n = 4, P value measured by Student’s unpaired t test). Scale bar = 50 μm. Results are expressed as mean ± SD; ** P < 0.01, *** P < 0.001, compared with PBS group; # P < 0.05, ## P < 0.01 compared with hUCMSC group; $ P < 0.05 compared with PBS group.

Article Snippet: PEDF and VEGF were quantified by PEDF ELISA kit (Elabscience, Wuhan, China) and VEGF ELISA kit (Proteintech).

Techniques: Transplantation Assay, Injection, Staining, TUNEL Assay, Immunostaining, In Vivo, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Effect of flavonoids hydroxygenkwanin on vascular smooth muscle cell proliferation, migration, and neointimal formation

doi: 10.1101/2022.12.20.521220

Figure Lengend Snippet: (A) The number of circulating blood EPCs was determined by flow cytometry 5 days after endothelial denudation (ED). Treatment with HGK (1 mg/kg) increased the number of circulating EPCs after ED. (B) Transwell assays showed that HGK promoted EPC migration. (C) Denuded femoral arteries were stained with Evans blue dye 7 days after ED. Femoral artery staining demonstrated that HGK promoted reendothelialization. (D) Immunohistochemical staining for CD31 was performed. HGK treatment increased CD31 expression (red) in denuded femoral arteries at 28 days after ED. Nuclei were stained with DAPI or hematoxylin. The thickness of neointimal hyperplasia is indicated by the double-headed arrows. The values are presented as the means ± SDs. *p <0.05 vs. the ED/CTRL or CTRL group. N=5-12. CTRL, control group. Statistical comparisons were performed using Student’s t test and one-way ANOVA.

Article Snippet: Human EPCs were obtained from CELLvo™ Human Endothelial Progenitor Cells (StemBioSys, San Antonio, TX) and then cultured in EGM-2 medium.

Techniques: Flow Cytometry, Migration, Staining, Immunohistochemical staining, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 1. Increased expression of PEDF in expanded human and mouse epidermis. A. Immunofluorescence staining for PEDF (green) and DAPI (blue) in the epithelial cells of human expanded skin (n = 9) and control skin (n = 6, ×40 magnification); B. Quantification of PEDF-positive cells per field in human skin (n = 6, mean ± SEM; **P < 0.01); C. Immunofluorescence staining for PEDF (green) and DAPI (blue) in the epithelial cells in mouse expanded skin and control skin (n = 6, ×40 magnification); D. Quantification of PEDF-positive cells per field in mouse skin (n = 6, mean ± SEM; ***P < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 3. PEDF accumulated in the subcutaneous exudates of a rat skin expansion model. The quantity of PEDF protein in the subcutaneous exudates obtained at indicated times in expanded and control rats were determined by ELISA assays (n = 6, mean ± SEM; ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 2. Skin expansion resulted in dermal thinning and PEDF up-regulation in a mouse skin expansion model. A. H&E staining at the 3rd week after expansion, showing thinner dermis in the expanded skin (×40 magnification); B. Quantification of dermal thickness between expanded and control skin (n = 6, mean ± SEM; **P < 0.01); C. The mRNA expression levels of PEDF in skin obtained at the 3rd week compared with the control skin (n = 6, mean

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Staining, Control, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 5. In vivo injection of PEDF promoted M1 macrophage polarization. A. A larger number of CD68+/iNOS+ double-positive macrophages were detected in the PEDF injection group compared with PBS treatment. In contrast to PEDF injection, LR antibody injection decreased the number of CD68+/iNOS+ double-positive macrophages; B. Quantification of M1 macrophages in differently treated skin (mean ± SEM; **P < 0.01，*P < 0.05).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: In Vivo, Injection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 4. Blockage of PEDF receptor rescued dermal thinning in vivo. A. HE staining results showed that recombinant PEDF protein injection decreased the thickness of expanded dermis, while LR blockage rescued dermal thinning of expanded skin; B. Quantification of dermal thickness between differently treated skin (n = 6, mean ± SEM; ***P < 0.001，*P < 0.05).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: In Vivo, Staining, Recombinant, Injection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 7. Hypoxia induced PEDF expression in HaCaT cells. The mRNA expression levels of PEDF in HaCaT cells under normoxic and hypoxic condi tions were determined by RT-qPCR (mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 8. PEDF promoted macrophages to polarize towards the M1 subtype under hypoxic conditions. A. FACs analysis of Raw264.7 single-cell sus pensions prepared after treating with PEDF under hypoxic conditions. CD11c+ cells indicate M1 macrophages. B. Quantification of CD11c+ cells under hypoxia (mean ± SEM; **P < 0.01). C. FACS analysis of Raw264.7 single-cell suspensions prepared after treating with PEDF under hypoxic conditions. CD206+ cells indicate M2 macro phages. D. Quantification of CD206+ cells under hypoxic conditions. The mRNA expression of M1 marker genes (E. iNOS, F. TNF-α) and M2 marker genes (G. Arg-1, H. Ym-1) determined by RT-qPCR and normalized to GAPDH mRNA expression. (mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Marker, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 9. PEDF hindered collagen syn thesis in a macrophage-mediated manner under hypoxic conditions. A, B. Relative mRNA expression of COLI (A) and COLIII (B) in fibroblasts after co-culture with PEDF treated macro phages determined by RT-qPCR (mean

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Co-Culture Assay, Quantitative RT-PCR