BacMam Virus Transduced Cardiomyoblasts Can Be Used for Myocardial Transplantation Using AP-PEG-A Microcapsules: Molecular Cloning, Preparation, andIn VitroAnalysis

Abstract

The potential of genetically modified cardiomyoblasts in treating damaged myocardium is well known. However, efficient delivery of these cells is of major concern during treatment. The limiting factors are the massive cell death that occurs soon after their intramyocardial transplantation into the beating heart. To address these problems, we generated recombinant baculoviruses (BacMam viruses) which efficiently transduced cardiomyoblast cells under optimized conditions. These genetically modified cells were then protected in a new polymeric microcapsule using poly-ethylene-glycol (PEG), alginate, and poly-L-lysine (PLL) polymers for efficient delivery. Results showed that microcapsules maintain cell viability and support cell proliferation for at least 30 days. The capsules exhibit strong immunoprotective potential and have high mechanical and osmotic stability with more than 70% intact capsules. The encased transduced cells showed a rapid transgene expression inside the capsule for at least 15 days. However, preclinical studies are needed to further explore its long-term functional benefits.

In Vitro and In Vivo Characterization of Temoporfin-Loaded Pegylated Plga Nanoparticles for Use in Photodynamic Therapy

“…In vitro analysis showed that Bac-NP-transduced cells were able to express their transgene for at least 3 weeks (Figure 2C), which is comparable to other experimental results. 24,31 This temporal expression of the delivery system is beneficial in many cases, particularly for angiogenesis where the expression of the therapeutic protein ceases once its job is done. Further studies need to be done to comprehend the exact mechanism of such a gene delivery system.…”

“…After 96 hours, absorbance was measured at 490 nm using CellTiter 96 ® AQueous Non-Radioactive Cell Proliferation Assay (Promega, Fitchburg, WI) in a Victor3 Multi Label Plate Counter (Perkin Elmer, Montreal, QC, Canada). 24 In a similar way, cardiomyocyte cell viabilities under oxidative stress in groups treated with different conditioned media were measured as described later in the study, using the same assay.…”

PAMAM Dendrimer-Baculovirus Nanocomplex for Microencapsulated Adipose Stem Cell-Gene Therapy: In Vitro and in Vivo Functional Assessment

“…The microencapsulated cells were cultured for 48 h in stationary flasks with standard medium, supplemented with antibiotics. The relative fluorescence units of each well containing 50 capsules resuspended in PBS were then measured using Victor3Multi Label Plate Counter (Perkin-Elmer, USA), as described elsewhere. , As shown in Figure A–D, the Bac MGFP -PAMAM (100) group showed significantly high normalized GFP expression (142.4 ± 16.8%) compared to Bac MGFP -PAMAM (10) (100 ± 7.5%) and Bac MGFP (33.6 ± 10.4%).…”

Angiopoietin-1-expressing adipose stem cells genetically modified with baculovirus nanocomplex: investigation in rat heart with acute infarction

“…After 96 hours, absorbance was measured at 490 nm using CellTiter 96 ® AQueous Non-Radioactive Cell Proliferation Assay (Promega, Fitchburg, WI) in a Victor3 Multi Label Plate Counter (Perkin Elmer, Montreal, QC, Canada). 24 In a similar way, cardiomyocyte cell viabilities under oxidative stress in groups treated with different conditioned media were measured as described later in the study, using the same assay. The experiment was performed in triplicate.…”