Cationic niosome-based hBMP7 gene transfection of neuronal precursor NT2 cells to reduce the migration of glioma cells in vitro

Abstract

This study explores interesting complementary approaches in cellbased hBMP7 gene delivery in order to mitigate the migration of glioblastoma cells based on the idea that this human bone morphogenetic protein (hBMP7) has enormous therapeutic potential in curing brain injuries as well as malignancies. After physicochemical characterization, the non-viral cationic niosomes were complexed with pUNO1-hBMP7 plasmids and used to transfect neuronal precursor NT2 cells. Subsequently, the transfected cells were co-cultured with glioma C6 cells to determine their antitumor effect in vitro. However the co-culture with either untransfected/transfected NT2 cells may reduce the viability of C6 glioma cells, the hBMP7-overexpressing NT2 cells hamper the migration of C6 glioma cells. These results highlight the potential of NT2 cell-based delivery of hBMP7 for impeding the metastasis of glioma cells. Fig. 1. Chemical structure of: A) Poloxamer 188 (P), B) Polysorbate 80 (P80), and C) Cationic lipid [2,3-di(tetradecyloxy)propan-1-amine] (D). 2. Material and methods 2.1. Material Human teratocarcinoma NTERA2/D1(NT2) (ATCC ® −CRL, 1973) and the rat glioma cell line (C6, CCL-107) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). C6 cells were grown in the ATCC-formulated F-12K Medium (Catalog No. 30-2004). Dulbecco's Modified Eagle's Medium (DMEM, ATCC 30 −2002), trypsin, Fetal bovine Serum (FBS) were purchased from Gibco ® (San Diego, California, US). Opti-MEM ® reduced medium, Lipofectamine® 2000 transfection reagent, antibiotics [100 U/ml penicillin and 100 μg/ ml streptomycin (Pen/Strep)] were acquired from Gibco® (Life Technologies S.A., Madrid, Spain). Cell counting (CCK-8) viability/ proliferation assay and crystal violet were obtained from Sigma Aldrich (Madrid, Spain). GelRed ® solution was obtained from Biotium (Hayward, California, USA), while other materials for gel electrophoresis were obtained from Bio-Rad (Madrid, Spain). pUNO1-hBMP7 plasmid (0.5 mg/ml) was obtained from InvivoGen (Toulouse, France). Sodium dodecyl sulfate (SDS), DNaseI, polysorbate 80, poloxamer 188, and PBS were purchased from Sigma Aldrich (Madrid, Spain). Uncoated Transwell ® 24 well microplates with 8.0 μm pore size (Costar-Corning, Corning, NY).

Niosome-Based Approach for In Situ Gene Delivery to Retina and Brain Cortex as Immune-Privileged Tissues

“…Therefore, two niosome formulations that differed only regarding the presence or absence of the non-ionic surfactant poloxamer 188 were elaborated by the reverse-phase evaporation technique and characterized to deliver the genetic material into the rat brain cortex. When poloxamer 188 was incorporated into the niosomes, the sizes of niosomes increased, which was probably due to the high HLB value of poloxamer 188 and to the interaction with the cationic lipid [161]. However, no significant change in zeta potential was reported, being over +40 mV.…”

Mesenchymal Stem Cells as a Gene Delivery Tool: Promise, Problems, and Prospects

“…Consequently, the closed bilayer structure of niosomes has hydrophilic inner and outer surfaces, with a sandwiched lipophilic area in between. The non-ionic surfactants are safe and affordable for the use in biomedicine as carriers for genes or hydrophilic/hydrophobic drugs [74]. Niosomes are prepared by various methods, such as ether injection, sonication, and microfluidics, to mention a few [75].…”

Cell-Based Therapy for the Treatment of Glioblastoma: An Update from Preclinical to Clinical Studies

“…Therefore, it was characterized by its ability to induce the differentiation of brain tumor stem cells. Attia et al [ 165 ] demonstrated human teratocarcinoma NTERA2/D1(NT2) cells transfected with hBMP7 nioplexes are a potential treatment vehicle for GB. The in vitro study exemplified the ability of BMP7-expressing neural precursor cells to lessen the tumorigenicity of glioma cells [ 165 ].…”