Stable Genetic Modification of Mesenchymal Stromal Cells Using Lentiviral Vectors

Martin, F. Elizabeth; Tristán‐Manzano, María; Maldonado‐Pérez, Noelia; Sánchez‐Hernández, Sabina; Benabdellah, Karim; Cobo, Manuel J.

doi:10.1007/978-1-4939-9065-8_17

Cited by 11 publications

(6 citation statements)

References 46 publications

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“…In the next step (Figure 4A Right and Figure 4B), which was aimed at demonstrating the cytoprotective properties of the long-term synergistic overexpression of the BCL2 and BDNF genes, the culture was continued for 12 days after the end of the transduction. As a result, a clear, statistically significant cytoprotective effect of the overexpression was demonstrated after using staurosporine to induce cell death by apoptosis via the intrinsic mitochondrial pathways (Figure 4B, staurosporine 0.5 µM and staurosporine 1 µM, EV vs. FV: p < 0.0001) [26]. In the case of doxorubicin (5 µM), we did not observe any differences between the EV and FV groups, so changes may have only been the effect of infection (Figure 4B, C vs. EV: p < 0.05), rather than a combination of Bcl-2 and BDNF overproduction (Figure 4B, EV vs. FV: ns).…”

Section: Cytoprotective Effect Depending On the Duration Of Bcl2 And Bdnf Overexpressionmentioning

confidence: 92%

Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

Borkowska

Zielińska

Paul-Samojedny

et al. 2021

IJMS

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Mesenchymal stem cells (MSC) are potentially a good material for transplantation in many diseases, including neurodegenerative diseases. The main problem with using them is the low percentage of surviving cells after the transplant procedure and the naturally poor ability of MSC to spontaneously differentiate into certain types of cells, which results in their poor integration with the host cells. The aim and the novelty of this work consists in the synergistic overexpression of two genes, BCL2 and BDNF, using lentiviral vectors. According to our hypothesis, the overexpression of the BCL2 gene is aimed at increasing the resistance of cells to stressors and toxic factors. In turn, the overexpression of the BDNF gene is suspected to direct the MSC into the neural differentiation pathway. As a result, it was shown that the overexpression of both genes and the overproduction of proteins is permanent and persists for at least 60 days. The synergistically transduced MSC were significantly more resistant to the action of staurosporine; 12 days after transduction, the synergistically transduced MSC had a six-times greater survival rate. The overexpression of the Bcl-2 and BDNF proteins was sufficient to stimulate a significant overexpression of the CHAT gene, and under specific conditions, the TH, TPH1, and SYP genes were also overexpressed. Modified MSC are able to differentiate into cholinergic and dopaminergic neurons, and the release of acetylcholine and dopamine may indicate their functionality.

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Section: Cytoprotective Effect Depending On the Duration Of Bcl2 And Bdnf Overexpressionmentioning

confidence: 92%

Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

Borkowska

Zielińska

Paul-Samojedny

et al. 2021

IJMS

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“…As the host micromilieu plays a crucial role in stem cell fate determination, the differentiation, maturation, and migration ability of hNS/PCs after transplantation is a decisive factor in determining their integration with host tissues [8]. Lentiviral gene transfer could be used as a promising tool to study the differentiation of hNS/PCs into various cell fates as well as to evaluate the cell migration into healthy host tissue in various pathological conditions, including SCI [18][19][20] Epilepsy surgery offers a unique opportunity to achieve hNS/PCs from the resected brain tissues of patients suffering from medically refractory epilepsy [21]. The hNS/PCs derived from epileptic brain tissues have the essential characteristics of neuroglial cells, which may provide an ideal source of autologous stem cell for replacement and restorative therapies [17,22,23].…”

Section: Introductionmentioning

confidence: 99%

Improvement of Rat Spinal Cord Injury Following Lentiviral Vector-Transduced Neural Stem/Progenitor Cells Derived from Human Epileptic Brain Tissue Transplantation with a Self-assembling Peptide Scaffold

et al. 2021

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Spinal cord injury (SCI) is a disabling neurological disorder that causes neural circuit dysfunction. Although various therapies have been applied to improve the neurological outcomes of SCI, little clinical progress has been achieved. Stem cell–based therapy aimed at restoring the lost cells and supporting micromilieu at the site of the injury has become a conceptually attractive option for tissue repair following SCI. Adult human neural stem/progenitor cells (hNS/PCs) were obtained from the epileptic human brain specimens. Induction of SCI was followed by the application of lentiviral vector-mediated green fluorescent protein–labeled hNS/PCs seeded in PuraMatrix peptide hydrogel (PM). The co-application of hNS/PCs and PM at the SCI injury site significantly enhanced cell survival and differentiation, reduced the lesion volume, and improved neurological functions compared to the control groups. Besides, the transplanted hNS/PCs seeded in PM revealed significantly higher migration abilities into the lesion site and the healthy host tissue as well as a greater differentiation into astrocytes and neurons in the vicinity of the lesion as well as in the host tissue. Our data suggest that the transplantation of hNS/PCs seeded in PM could be a promising approach to restore the damaged tissues and improve neurological functions after SCI.

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“…Retroviruses can integrate efficiently into the chromatin of target cells, which is ideal for increased efficiency [5]. Lentiviruses' strategy is to increase the transfection rate as they rely on the machinery of the cell and can target non-dividing cells [11,17]. Finally, adeno-associated viruses have been mainly utilized for genome transduction, which targets both dividing and non-dividing cells [11,18].…”

Section: Introductionmentioning

confidence: 99%

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

et al. 2020

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Over the past decade, gene therapies have attracted much attention for the development of treatments for various conditions, including cancer, neurodegenerative diseases, protein deficiencies, and autoimmune disorders. Despite the benefits of this approach, several challenges are yet to be solved to reach clinical implementation. Some of these challenges include low transfection rates, limited stability under physiological conditions, and low specificity towards the target cells. An avenue to overcome such issues is to deliver the therapies with the aid of potent cell-penetrating vectors. Non-viral vectors, such as nanostructured materials, have been successfully tested in drug and gene delivery. Here, we propose the development and in vitro evaluation of a nanostructured cell-penetrating vehicle based on core/shell, magnetite/silver nanoparticles. A subsequent conjugation of a pH-responsive polymer was used to assure that the vehicle can carry and release circular DNA. Additionally, the translocating peptide Buforin II was conjugated with the aid of a polyether amine polymer to facilitate translocation and endosome escape. The obtained nanobioconjugates (magnetite/silver-pDMAEMA-PEA-BUFII) were characterized by UV-Vis spectrophotometry, dynamic light scattering (DLS), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), scanning electron microscope equipped with energy dispersive spectroscopy (SEM+EDS), and transmission electron microscopy (TEM). They were also encapsulated in lecithin liposomes to form magnetoliposomes. The cell viability of Vero cells in the presence of the nanobioconjugates was above 95% and declined to 80% for the magnetoliposomes. The hemolytic tendency of nanobioconjugates and magnetoliposomes was below 10%, while the platelet aggregation approached that of the negative control (i.e., 35%). Cytoplasm coverage values of about 50% for both Vero and neuroblastoma cells confirmed significant cell penetration. Pearson’s correlation coefficients for both cell lines allowed us to estimate 20–40% colocalization of the nanobioconjugates with lysotracker green, which implied high levels of endosomal escape. The developed vehicles were also capable of loading around 16% of the added DNA and releasing such cargo with 8% efficiency. The developed nanoplatform holds a significant promise to enable highly efficient gene therapies as it overcomes some of the major issues associated with their eventual translation to the pre-clinical and clinical scale.

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Stable Genetic Modification of Mesenchymal Stromal Cells Using Lentiviral Vectors

Cited by 11 publications

References 46 publications

Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

Improvement of Rat Spinal Cord Injury Following Lentiviral Vector-Transduced Neural Stem/Progenitor Cells Derived from Human Epileptic Brain Tissue Transplantation with a Self-assembling Peptide Scaffold

PH-Responsive, Cell-Penetrating, Core/Shell Magnetite/Silver Nanoparticles for the Delivery of Plasmids: Preparation, Characterization, and Preliminary In Vitro Evaluation

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