K2 Transfection System Boosts the Adenoviral Transduction of Murine Mesenchymal Stromal Cells

Abstract

Adenoviral vectors are important vehicles for delivering therapeutic genes into mammalian cells. However, the yield of the adenoviral transduction of murine mesenchymal stromal cells (MSC) is low. Here, we aimed to improve the adenoviral transduction efficiency of bone marrow-derived MSC. Our data showed that among all the potential transduction boosters that we tested, the K2 Transfection System (K2TS) greatly increased the transduction efficiency. After optimization of both K2TS components, the yield of the adenoviral transduction increased from 18% to 96% for non-obese diabetic (NOD)-derived MSC, from 30% to 86% for C57BL/6-derived MSC, and from 0.6% to 63% for BALB/c-derived MSC, when 250 transduction units/cell were used. We found that MSC derived from these mouse strains expressed different levels of the coxsackievirus and adenovirus receptors (MSC from C57BL/6≥NOD>>>BALB/c). K2TS did not increase the level of the receptor expression, but desensitized the cells to foreign DNA and facilitated the virus entry into the cell. The expression of Stem cells antigen-1 (Sca-1) and 5′-nucleotidase (CD73) MSC markers, the adipogenic and osteogenic differentiation potential, and the immunosuppressive capacity were preserved after the adenoviral transduction of MSC in the presence of the K2TS. In conclusion, K2TS significantly enhanced the adenoviral transduction of MSC, without interfering with their main characteristics and properties.

Treatment with Mesenchymal Stromal Cells Overexpressing Fas-Ligand Ameliorates Acute Graft-versus-Host Disease in Mice

“…Third, although mRNA expression of FasL was detected 8 h following adenoviral transduction [ 25 ], detection of the GFP reporter protein was highly variable. GFP expression in BALB/c-derived MSC was detected when the cells were transduced with more than 500 TU/cell [ 28 ]. Thus, FasL expression exceeded GFP almost eight times compared to cells transduced with the empty vector control adenovirus.…”

Transduction Enhancers Enable Efficient Human Adenovirus Type 5-Mediated Gene Transfer into Human Multipotent Mesenchymal Stromal Cells

“…The transduction-enhancing effect of cationic polymers is in line with the results reported by other researchers. Not only polybrene [16] and poly-L-lysine [18,20], but also polyethylenimine (PEI) [46], the K2 transfection system [18], and lipofectamine [47][48][49] are cationic polymers and lipids known to enhance HAdV-5-mediated gene transfer in different cell types. As all these molecules share their cationic charge, it is hypothesized that these polymers and lipids form complexes with the negatively charged adenoviral particles, which can then efficiently enter the cells.…”

“…Several cationic polymers and lipids, including polybrene [16], poly-L-lysine [17] or the K2 transfection system [18], were reported to increase the HAdV-5 transduction efficiency in various cell types, including hMSCs [17,19,20]. Besides, some proteins are known to interact with the HAdV-5 capsid, thereupon mediating cell transduction, both in vivo and in vitro [21][22][23][24].…”