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Overview of the experimental workflow. The workflow illustrates key steps, including UC-MSCs isolation, plasmid construction, gene transfection and evaluation, secretome production, BMP-2 mRNA and protein quantifications, and secretome-BMP-2 functional assay in osteogenic differentiation of untransfected cells. (Created in BioRender. Fahdia, N. (2026). https://BioRender.com/2bdayt7 ).

Journal: Biomedicines

Article Title: Genetic Engineering of Umbilical Cord-Derived Mesenchymal Stem Cells to Enhance BMP-2 Secretion via Signal Peptide Optimization

doi: 10.3390/biomedicines14010076

Figure Lengend Snippet: Overview of the experimental workflow. The workflow illustrates key steps, including UC-MSCs isolation, plasmid construction, gene transfection and evaluation, secretome production, BMP-2 mRNA and protein quantifications, and secretome-BMP-2 functional assay in osteogenic differentiation of untransfected cells. (Created in BioRender. Fahdia, N. (2026). https://BioRender.com/2bdayt7 ).

Article Snippet: Quantification of BMP-2 protein in the secretome was performed using a human BMP-2 ELISA kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol.

Techniques: Isolation, Plasmid Preparation, Transfection, Functional Assay

Design of the plasmid incorporating a signal peptide in the BMP-2 sequence. The SP sequence is located at the N-terminus of the BMP-2 amino acid sequence and consists of the N-, H-, and C-regions. The SP sequence is directly fused to the BMP-2 sequence to facilitate efficient targeting into the secretory pathway.

Journal: Biomedicines

Article Title: Genetic Engineering of Umbilical Cord-Derived Mesenchymal Stem Cells to Enhance BMP-2 Secretion via Signal Peptide Optimization

doi: 10.3390/biomedicines14010076

Figure Lengend Snippet: Design of the plasmid incorporating a signal peptide in the BMP-2 sequence. The SP sequence is located at the N-terminus of the BMP-2 amino acid sequence and consists of the N-, H-, and C-regions. The SP sequence is directly fused to the BMP-2 sequence to facilitate efficient targeting into the secretory pathway.

Article Snippet: Quantification of BMP-2 protein in the secretome was performed using a human BMP-2 ELISA kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol.

Techniques: Plasmid Preparation, Sequencing

Evaluation of transfection efficiency, cell viability, and BMP-2 secretion in UC-MSCs transfected with different signal peptide constructs: ( A ) Representative brightfield and fluorescence microscopy images showing GFP expression. ( B ) Representative flow cytometry dot plots showing GFP-negative (untransfected) and GFP-positive (transfected) UC-MSC populations. ( C ) Quantitative analysis of GFP expression (%). ( D ) Cell viability (%) after transfection. ( E ) Relative BMP2 mRNA expression quantified by qRT-PCR and ( F ) BMP-2 protein secretion quantified by ELISA at 48 h post-transfection. ( G ) BMP-2 protein levels in UC-MSC secretome across serial passages. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test; p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Journal: Biomedicines

Article Title: Genetic Engineering of Umbilical Cord-Derived Mesenchymal Stem Cells to Enhance BMP-2 Secretion via Signal Peptide Optimization

doi: 10.3390/biomedicines14010076

Figure Lengend Snippet: Evaluation of transfection efficiency, cell viability, and BMP-2 secretion in UC-MSCs transfected with different signal peptide constructs: ( A ) Representative brightfield and fluorescence microscopy images showing GFP expression. ( B ) Representative flow cytometry dot plots showing GFP-negative (untransfected) and GFP-positive (transfected) UC-MSC populations. ( C ) Quantitative analysis of GFP expression (%). ( D ) Cell viability (%) after transfection. ( E ) Relative BMP2 mRNA expression quantified by qRT-PCR and ( F ) BMP-2 protein secretion quantified by ELISA at 48 h post-transfection. ( G ) BMP-2 protein levels in UC-MSC secretome across serial passages. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test; p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Article Snippet: Quantification of BMP-2 protein in the secretome was performed using a human BMP-2 ELISA kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol.

Techniques: Transfection, Construct, Fluorescence, Microscopy, Expressing, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay