We aim to identify a suitable method for long-term monitoring of the migration and proliferation of mesenchymal stromal cells in a rat model of stroke using transgene expression of ferritin by magnetic resonance imaging (MRI). bone marrow mesenchymal stromal cells (BMSCs) were transduced with lentivirus containing plasmid shuttle (pCDH-CMV-MCS-EF1-copGFP) brings ferritin heavy chain 1 (Fth1) gene. Ferritin expression in stromal cells was evaluated by western blotting and immunofluorescent staining.
Absorption of iron from Fth1-BMSCs measured by Prussian blue staining. Following the introduction of surgical occlusion of the middle cerebral artery, Fth1-BMSCs and superparamagnetic iron oxide- (SPIO-) labeled BMSCs were injected via the internal jugular vein. Imaging and the intensity of the signal being monitored by diffusion-weighted imaging (DWI), T2-weighted imaging (T2WI), and susceptibility-weighted imaging (SWI) in vitro and in vivo. Pathology conducted for comparison. We observed that the MRI signal intensity of SPIO-BMSCs gradually reduced over time.
Fth1-BMSCs showed the same signal intensity between 10 and 60 days. SWI shows hypointense lesions in SPIO-BMSC (traceable to 30 d) and Fth1-BMSC group. T2WI not sensitive enough to track Fth1-BMSCs. After transplantation, the cells stained with Prussian blue was observed around the central area of infarction and infarction in both transplantation models. Fth1-BMSCs transplanted to treat focal cerebral infarction safe, reliable, and can be tracked by MRI. Fth1 more stable and suitable labeling of SPIO labeling for long-term tracking. SWI is more sensitive than T2W1 and suitable as MRI-tracking optimal sequence.
MRI Tracking of SPIO- and Fth1-Labeled Bone Marrow Mesenchymal Stromal Cell Transplantation for Treatment of Stroke.
Design and analysis of integrated reporter’s stable transgene expression is induced in human T cells and NK-cell line CAR.
the cytotoxic activity of T and NK-cells can be efficiently Retargeted against cancer cells using chimeric antigen receptor (CAR) and rTCRs. In the context of solid cancers, the use of armored cells CAR T and NK secrete a molecule anti-cancer extras such as cytokines, chemokines, antibodies, bite, cytokine receptor inverse, and inhibitors of the checkpoint, appears particularly promising, as this can help to overcome the immunosuppressive tumor micro , attracting an audience of immune cells, and increasing the CAR T / NK-cell persistence.
Placing such molecule expression under the control of the downstream transcription activation offers T / NK-cell-mediated CAR advantages of targeted delivery, high local concentrations and reduced toxicity. Some canonical DNA sequences are known to function as a promoter-induced activation in human T and B cells have been described to date and usually include multimers of NFkB and NFAT binding sites. However, relatively little is known about the sequence of DNA that can serve as a switch in a context-driven activation of NK cells.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: The ANPRA/Aequorin expression vector is designed to co-express human atrial natriuretic peptide receptor A (ANPRA, also called natriuretic peptide receptor A/guanylate cyclase A) and jellyfish (Aequorea victoria) Aequorin in mammalian cells.
Description: GFP-Rac1 Expression Vector Set contains 3 vectors: Rac wild type, T17N dominant negative mutant, and Q61L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: GFP-RhoA Expression Vector Set contains 3 vectors: RhoA wild type, T19N dominant negative mutant, and Q63L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: GFP-Cdc42 Expression Vector Set contains 3 vectors: Cdc42 wild type, T17N dominant negative mutant, and Q61L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: Active Rac1 Expression Vector Set contains 3 vectors expressing different constitutively active mutants of Rac1: Q61L, Q61L/F37A, and Q61L/Y40C.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Active H-Ras Expression Vector Set contains 3 vectors expressing different constitutively active mutants of H-Ras: V12, V12S35, and V12C40.
Description: Baculovirus cassette vector pAc-l-CH3 for the expression of human, humanized or chimeric IgG(lambda) in insect cells and secretion of assembled antibodies into the supernatant.
We set out to compare the functionality of some activation-induced promoter in primary human T cells, as well as in the line of NK cells and NK-92 construction YT.Lentiviral engineered to express two fluorescent reporters: mCherry under 4xNFAT, 2xNFkB, 5xNFkB, 10xNFkB, 30xNFkB promoter, as well as two promoter variants of CD69, and copGFP under a strong constitutive promoter of the gene of human EF1a. Pseudotyped lentiviral particles obtained using the construction transduced into human primary T cells and NK-92 and certain cell lines expressing YT CAR to PSMA. Transgenic cells derived activated by CD3 / CD28 beads (T cell) or via CAR (CAR-NK cell line).
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