fibroblast clones  (Trans Ova)

Journal: bioRxiv

Article Title: Cytomegalovirus pentamer dependent endocytic cellular infection utilizes redundant entry receptors in the guinea pig model

doi: 10.64898/2026.05.07.723586

Figure Lengend Snippet: (A-B) Fibroblast GPL cells. (A) GPCMV(PC+) growth (MOI 1 pfu/cell, 3 DPI) on PDGFRA/NRP2 DKO GPL cells (black) compared to DKO cells ectopically expressing either NRP2 (green) or ThBD (red). Control wild type GPL(blue). One-way ANOVA p < 0.05. (B) GPCMV(PC+) growth on PDGFRA/NRP2 DKO GPL (black) compared to PDGFRA/NRP2 DKO GPL + ectopic expression of CD46 (orange). Both cell lines had minimal support for GPCMV infection. Student t test, ns = not significant). (C-D) Epithelial REPI cells. (D) GPCMV(PC+) growth (MOI 1 pfu/cell, 3DPI) on ThBD/NRP2 DKO REPI cells or DKO cells ectopically expressing NRP2 (green) or ThBD (red). Control wild type REPI (blue). One-way ANOVA p < 0.05. (D) GPCMV(PC+) growth on ThBD/NRP2 DKO GPL (black) compared to PDGFRA/NRP2 DKO GPL + ectopic expression of CD46 (orange). Student t test, ns = not significant).

Article Snippet: However, studies from the mid-1990s onwards utilized an established ATCC embryo fibroblast cell line (GPL ATCC CCL 158) for virus growth and titration experiments because of an ability of cells to be contact inhibited as a confluent monolayer and these cells easily supports virus, BAC and plasmid transfections ( ).

Techniques: Expressing, Control, Infection

Journal: bioRxiv

Article Title: Cytomegalovirus pentamer dependent endocytic cellular infection utilizes redundant entry receptors in the guinea pig model

doi: 10.64898/2026.05.07.723586

Figure Lengend Snippet: Wild type GPCMV virus stock generated on REPI cells (REPI stock) or by single passage on fibroblast GPL cells (GPL stock) were evaluated for ability to infect GPL fibroblasts, REPI and trophoblast (TEPI) epithelial cells. (A) Comparative one-step growth curve of GPCMV (MOI 1 pfu/cell) on GPL and PDGFRA KO GPL cells. Viral titers evaluated for 1-6 days post infection (dpi). GPCMV GPL stock (triangle), GPL cell infection (blue), PDGFRA KO GPL cells (black). GPCMV REPI stock (square), GPL cell infection (green), PDGFRA KO GPL cells (green). (B) Comparative one-step growth curve of GPCMV on REPI or TEPI cells by virus stock generated on GPL cells (triangle) or REPI cells (square). Experimental set up similar to (A) with MOI of 1 pfu/cell. GPCMV (GPL stock) infection of REPI cells (green triangle), or TEPI cells (blue triangle). GPCMV (REPI stock) infection of REPI cells (grey square), or TEPI cells (purple square). Viral titers (1-6 dpi) were determined on GPL cells as described for A. Results shown are average values from replicates carried out in triplicate.

Article Snippet: However, studies from the mid-1990s onwards utilized an established ATCC embryo fibroblast cell line (GPL ATCC CCL 158) for virus growth and titration experiments because of an ability of cells to be contact inhibited as a confluent monolayer and these cells easily supports virus, BAC and plasmid transfections ( ).

Techniques: Virus, Generated, Infection

Journal: Molecular Therapy Advances

Article Title: Swine reporter model for preclinical evaluation and characterization of gene delivery vectors

doi: 10.1016/j.omta.2026.201729

Figure Lengend Snippet: Generation of the SRM-1 Cre- or CRISPR-inducible swine reporter model (A) Schematic illustration of the SRM-1 reporter integrated into the swine ROSA26 gene. Reporter design includes loxP sites (purple triangles) and CRISPR sites (blue and red arrows) for Cre- or genome editing-mediated reporter activation, respectively. (B) PCR of the integration junction of the SRM-1 construct in the ROSA26 gene and SRM-1 construct copy number measured by ddPCR in founder SRM-1 animals. Asterisk indicates an animal with imprecise reporter integration. (C) The percentage of tdTomato-positive SRM-1 fibroblasts measured by flow cytometry after transfection with either Cre mRNA, SpCas9 RNP, or AsCas12a RNP. Open circles are individual data points; error bars are standard error of the mean. (D) Schematic of the probe-based assay developed to measure SRM-1 DNA recombination by Cre. Without Cre-induced DNA recombination of the loxP sites, the SacI restriction enzyme will cut the DNA and not allow for PCR from the forward and reverse primers (black arrows). After DNA recombination, the SacI site is removed from the amplicon and PCR will cause degradation of the FAM-conjugated probe (gray arrow) which will release the FAM fluorescence (green arrow). (E) Comparison of SRM-1 reporter activation by tdTomato expression measured by flow cytometry and by DNA recombination measured by ddPCR across a 100-fold drop in Cre mRNA transfection. Open circles are individual measurements and bars are the average measurement for tdTomato (black) and ddPCR (purple). Brackets with numbers are the difference in percentages between the average flow cytometry measurement and the average ddPCR measurement.

Article Snippet: Fibroblast clones were shipped to TransOva Genetics for nuclear transfer using their established protocols.

Techniques: CRISPR, Activation Assay, Construct, Flow Cytometry, Transfection, Amplification, Fluorescence, Comparison, Expressing

Journal: ACS Omega

Article Title: Comparative Study of Chitosan-Pyrophosphate and Magnesium Hydroxide-Alginate Hybrid Nanoparticles: Physicochemical Properties and Cytocompatibility toward Vascular Calcification Applications

doi: 10.1021/acsomega.5c12883

Figure Lengend Snippet: (a) Cytotoxicity analysis of L929 cells after 24 h of exposure to NPs-Mg­(OH) 2 -Alg at different concentrations. * p < 0.05 vs control; (b) cytotoxicity analysis of L929 cells after 24 h of exposure to NPs-CS-PPi at different concentrations. * p < 0.05 vs control.

Article Snippet: The murine fibroblast cell line L929 (ATCC NCTC clone 929 [L cell, CCL-1]) was used for cytotoxicity assays.

Techniques: Control