human rpe cell line arpe19  (Procell Inc)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Retinal Pigment Epithelium Extracellular Vesicles Induce Microglia Polarization in MERTK -Associated Retinal Degeneration

doi: 10.1167/iovs.66.15.1

Figure Lengend Snippet: Establishment and characterization of in vitro RPE research model with RP-associated MERTK gene mutation. ( A ) Workflow of patient-specific iPSC and iPSC-RPE generation. ( B ) Fundus photography, fundus autofluorescence imaging, and OCT of the RP patient with the MERTK mutation. ( C ) Fundus photography, fundus autofluorescence imaging, and OCT of a healthy control. ( D ) Phase-contrast images ( left ) and SEM imaging ( right ) of iPSC-RPE from the control. Scale bars : 100 µm ( D1 , D2 ), 5 µm ( D3 ). ( E ) Phase-contrast images ( left ) and SEM imaging ( right ) of iPSC-RPE from an RP patient. Patient iPSC-RPE cells also formed a tightly packed monolayer across the culture plates, except for local rosette-like cell clusters. Scale bars : 100 µm ( E1 , E2 ), 5 µm ( E3 ). ( F ) Western blot analysis of MERTK protein expression in the ARPE19 line, control, and patient in the in vitro RPE model. ( G, H ) Immunofluorescence staining of MERTK and its phosphorylated protein PAXL in control iPSC-RPE ( G ) and patient iPSC-RPE ( H ). Scale bars : 20 µm.

Article Snippet: Because the lack of MERTK protein expression was observed in the human RPE cell line ARPE19 obtained from the Procell Life Science & Technology Co., Ltd. (Wuhan, China), we utilized iPSC technology to establish a stable patient-specific in vitro research model of RPE.

Techniques: In Vitro, Mutagenesis, Imaging, Control, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: Rescue of TMEM97 expression in TMEM97 KO ARPE19 cells hampers oxidant-induced ROS elevation ARPE19 cell culture and ROS detection are described in . Three independent cell cultures ( n = 3) were used, and 25 slides were imaged and averaged to generate the mean ± SEM. Statistics: two-way ANOVA; ∗∗∗ p < 0.001 (compared to the respective WT control); # p < 0.05; ### p < 0.001 (pairwise comparison).

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Expressing, Cell Culture, Control, Comparison

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: Proteomics analysis of the impact of TMEM97 ablation on protein expression in ARPE19 cells TMEM97 +/ + (WT) and TMEM97 −/ − (KO) ARPE19 cells were cultured to full confluence in the regular growth medium. For proteomic analysis, 4 conditions were applied: WT and KO cells were treated without or with 5 mM NaIO 3 for 24 h before harvest for mass spectrometry. For transcriptomic analysis, WT and KO cells were cultured without NaIO 3 . Three independent cultures ( n = 3) of WT or KO cells were used. (A) Proteomics heatmap. Top 10 best q values ( p adj, adjusted p value) for upregulated proteins (KO vs. WT) were selected, and the proteins were ranked by their fold changes; n = 3. (B) GO enrichment. Presented are the top 20 upregulated pathways ranked by best q values ( p adj). (C) Volcano plots. Red and green dots represent up- and downregulated proteins, respectively. The dashed line is a threshold of p = 0.05. (D) Transcriptomics heatmap. Top 10 best q values ( p adj) for upregulated genes (KO vs. WT) were selected, and the genes were ranked by their expression fold changes; n = 3.

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Expressing, Cell Culture, Mass Spectrometry

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: Immunoblots indicate that TMEM97 suppresses E-cadherin and N-cadherin protein expression To align WT and TMEM97 KO ARPE19 cells to an epithelial cell state, the cells were cultured to full confluence and then maintained for 3 days prior to harvest for immunoblot assays. For the rescue of TMEM97 expression, lentivirus for the EV control or for TMEM97 OE was used to transduce WT and KO cells that reached full confluence and incubated for 3 days prior to harvest for immunoblotting. Quantification: mean ± SEM, n = 3–8 independent repeat experiments (indicated by data points in A and B). Student’s t test (2 groups) or one-way ANOVA/Tukey (4 groups): ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001 (compared to the first bar); #### p < 0.0001 (pairwise comparison). (A) TMEM97 negates the protein levels of AXL, ADAM10, and POSTN. (B) TMEM97 ablation has no effect on α-SMA, vimentin, fibronectin, and β-catenin protein levels but does result in upregulation of E-cadherin and N-cadherin proteins. (C) TMEM97 ablation increases the ZO-1 protein level. For the immunofluorescence assay, ARPE19 cells were cultured to subconfluence. Quantification: For WT or KO cells, 64 images were taken, and the fluorescence intensities of 16 areas within each image were measured using ImageJ and averaged; the mean ± SEM of each group was then calculated. Statistics: Student’s t test; ∗∗∗ p < 0.001.

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Western Blot, Expressing, Cell Culture, Control, Transduction, Incubation, Comparison, Immunofluorescence, Fluorescence

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: Immunofluorescence indicates upregulation of E- and N-cadherins in TMEM97 KO ARPE19 cells To align WT and TMEM97 KO ARPE19 cells to an epithelial cell state, the cells were cultured to full confluence and then maintained for 3 days prior to assays. For the rescue of TMEM97 expression, WT and KO cells that reached full confluence were added with the lentivirus for the EV control or for TMEM97 OE and cultured for 3 days prior to harvest for immunoblot or immunofluorescence. (A) Immunoblots of E-cadherin and N-cadherin. Quantification: mean ± SEM, n = 3–4 independent repeat experiments. Statistics: one-way ANOVA/Tukey; ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001 (compared to the first bar, EV/WT); #### p < 0.0001 (pairwise comparison) . (B) Immunofluorescence of E-cadherin (red) and ZO-1 (green). (C) Immunofluorescence of N-cadherin (red) and ZO-1 (green).

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Immunofluorescence, Cell Culture, Expressing, Control, Western Blot, Comparison

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: TMEM97 ablation induces ARPE19 cell morphologic change and proliferation To capture changes in cell growth and morphology, WT and TMEM97 KO ARPE19 cells cultured to ∼50% confluence were used for various analyses. For the rescue of TMEM97 expression, lentivirus for the EV control or for TMEM97 OE was used to transduce WT and KO cells for 3 days before harvest for assays. (A) Representative images of cell morphometric segmentation. (B) Schematic of 3 categories of cell circularity. Green images are representative calcein-stained ARPE19 cells. (C) Bin plot showing quantification of cell circularity. Percentages versus total cell numbers are provided for each category. Statistics: Student’s t test; ∗∗∗∗ p < 0.0001. (D) Immunoblots. Quantification: mean ± SEM, n = 3–4 independent repeat experiments. Student’s t test (2 groups) or one-way ANOVA/Tukey (4 groups); ∗∗∗∗ p < 0.0001 (compared to the first bar); #### p < 0.0001 (pairwise comparison). (E) mRNA levels determined by qRT-PCR. In this experiment, OE was conducted using a single-clone cell line generated through lentiviral transduction of ARPE19 cells. Quantification: mean ± SD, n = 3 repeats. Student’s t test: ∗∗ p < 0.01; ∗∗∗ p < 0.001. (F) Cell growth. Cells were stained with the fluorescent dye calcein, and the cell numbers were quantified (each normalized with the cell number at the beginning of the culture). ARPE19∗, the ARPE19 cells without lentiviral transduction. Quantification: Mean ± SD, n = 8. One-way ANOVA/Tukey: ∗∗∗∗ p < 0.0001 (compared to ARPE19∗); ### p < 0.001 (pair-wise comparison). (G) Summary of TMEM97 KO-induced changes in epithelial and mesenchymal marker expression. ∗∗Claudin levels based on the transcriptomics data. ↑, upregulated; ↓, downregulated; ↔, no change.

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Cell Culture, Expressing, Control, Transduction, Staining, Western Blot, Comparison, Quantitative RT-PCR, Generated, Marker

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: TMEM97 ablation induces ARPE19 cell clustering (A) Clustering of cultured ARPE19 cells. To observe clustering, WT and TMEM97 KO cells were cultured to ∼50% confluence and calcein stained. Three representative images are presented for WT or KO cells. (B) Schematic indicating the mouse model of oxidant-induced RPE damage. Mice received a single tail-vein injection of NaIO 3 (30 mg/kg) and were euthanized 3 days later. (C) Immunofluorescence of ZO-1 on retinal cryosections. For each animal group, the images were randomly picked from that of 3 mice. Day 0 represents the control group injected with PBS. OLM, outer limiting membrane. Red arrow indicates the RPE layer; ∗ clustering-like RPE cell aggregates. Scale bar: 20 μm.

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Cell Culture, Staining, Injection, Immunofluorescence, Control, Membrane

Journal: Molecular Therapy. Nucleic Acids

Article Title: TMEM97 governs partial epithelial-mesenchymal transition of retinal pigment epithelial cells via the CTNND2-ADAM10 axis

doi: 10.1016/j.omtn.2025.102460

Figure Lengend Snippet: TMEM97 loss and gain of function reveal a CTNND2-ADAM10 axis that supports E- and N-cadherin protein levels To align WT and TMEM97 KO ARPE19 cells to an epithelial cell state, the cells were cultured to full confluence and then maintained for 3 days prior to harvest for immunoblot assays. For the rescue of TMEM97 expression, lentivirus for the EV control or for TMEM97 OE was used to transduce WT and KO cells that reached full confluence. The transduction was maintained for 3 days prior to harvest for immunoblotting. Quantification: mean ± SEM, n = 3–4 independent repeat experiments. Student’s t test (2 groups) or one-way ANOVA/Tukey (4 groups): ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001 (compared to the first bar); # p < 0.05; ### p < 0.001; #### p < 0.0001 (pairwise comparison). (A) AXL regulates E-cadherin but not N-cadherin protein levels. (B) ADAM10 regulates both E- and N-cadherin protein levels. (C) TMEM97 negatively regulates CTNND2 protein levels. (D) CTNND2 knockdown affects ADAM10 but not AXL protein levels. (E) CTNND2 knockdown reduces both E- and N-cadherin protein levels. (F) Schematic of the TMEM97-CTNND2-ADAM10 pathway that regulates the concurrent E- and N-cadherin expression.

Article Snippet: The ARPE19 human RPE cell line was from the American Type Culture Collection (catalog no. CRL2302).

Techniques: Cell Culture, Western Blot, Expressing, Control, Transduction, Comparison, Knockdown