Polyethylenimine600-β-cyclodextrin: a promising nanopolymer for nonviral gene delivery of primary mesenchymal stem cells

Abstract

Genetically modified mesenchymal stem cells (MSCs) have great potential in the application of regenerative medicine and molecular therapy. In the present manuscript, we introduce a nanopolymer, polyethylenimine 600 -β-cyclodextrin (PEI 600 -β-CyD), as an efficient polyplex-forming plasmid delivery agent with low toxicity and ideal transfection efficiency on primary MSCs. PEI 600 -β-CyD causes significantly less cytotoxicity and apoptosis on MSCs than 25 kDa high-molecular-weight PEI (PEI25kDa). PEI 600 -β-CyD also exhibits similar transfection efficiency as PEI25kDa on MSCs, which is higher than that of PEI600Da. Quantum dot-labeled plasmids show that PEI 600 -β-CyD or PEI25kDa delivers the plasmids in a more scattered manner than PEI600Da does. Further study shows that PEI 600 -β-CyD and PEI25kDa are more capable of delivering plasmids into the cell lysosome and nucleus than PEI600Da, which correlates well with the results of their transfection-efficiency assay. Moreover, among the three vectors, PEI 600 -β-CyD has the most capacity of enhancing the alkaline phosphatase activity of MSCs by transfecting bone morphogenetic protein 2, 7, or special AT-rich sequence-binding protein 2. These results clearly indicate that PEI 600 -β-CyD is a safe and effective candidate for the nonviral gene delivery of MSCs because of its ideal inclusion ability and proton sponge effect, and the application of this nanopolymer warrants further investigation.

Development of a Gene-Activated Scaffold Incorporating Multifunctional Cell-Penetrating Peptides for pSDF-1α Delivery for Enhanced Angiogenesis in Tissue Engineering Applications

“…Interestingly, GET* N/P 8 nanoparticles elicited the highest transfection efficiency in MSCs in 2D, significantly more so than unmodified GET and GET* N/P 6, and was also higher than the GET* N/P 11 group. This contrasts with similar optimisation studies in MSCs, where increasing N/P ratio tends to correlate with increased transfection efficiency [58,59]. One potential hypothesis to explain this result is that the increased positive charge conferred on GET* N/P 11 particles as a result of an increased proportion of FLH to FLR causes a more pronounced 'proton-sponge' effect and subsequently less viable cells can be transfected.…”

miR‐33a‐5p modulates TNF‐α‐inhibited osteogenic differentiation by targeting SATB2 expression in hBMSCs

“…hBMSCs were subjected to lineage identification according to a previously reported protocol . The antibodies used included anti‐CD‐45 (#340953; BD Biosciences, San Jose, CA, USA), anti‐CD34 (#551401; BD Pharmingen, San Jose, CA, USA), anti‐CD44 (#555478; BD Pharmingen), anti‐CD90 (#551401; BD Pharmingen), anti‐CD105 (#323208; Biolegend, San Diego, CA, USA), and anti‐CD29 (#555443; BD Pharmingen).…”

Low-dose Polyethylenimine-Mediated Transfection to Generate Human Induced Pluripotent Stem Cell: an optimized protocol

“…In the current investigation, we represented a cost-effective and optimized protocol for transfecting HDFs through a gold standard reagent (i.e PEI) and oriP/EBNA1-based vectors to produce hiPSCs. Previous evidences have con rmed the capability of PEI to transfect several types of cells such as HEK293 [2], mesenchymal [39], neural [40], and embryonic [41] stem cells. However, with the aim of generating hiPSCs, our investigation is of the pioneers setting and standardizing several parameters relevant to HDF transfection process using PEI25K.…”