The immunomodulatory and self-renewable features of human adipose mesenchymal stem cells (HAD-MSCs) mark their importance in regenerative medicine. Interleukin 23 (IL-23) as proinflammatory cytokines Hitting the regulatory T cells (Treg) and promote the response of T helper 17 (Th17) and helper 1 (Th1) cells T. This path begins inflammation and immunosuppression in several autoimmune diseases. The present study was to produce recombinant IL-23 decoy receptor (RIL- 23R) using own-MSCs as a good candidate for the former purpose vivo cell-based gene therapy reduces inflammation in autoimmune disease. have-MSCs isolated from lipoaspirate and then marked by differentiation.
RIL- 23R is designed and cloned into pCDH-813A- 1 lentiviral vectors. Transduction-MSCs have done in MOI (multiplicity of infection) = 50 with EFI pCDH- α- RIL- 23R- copGFP PGK. RIL- 23R expression and octamer-binding transcription factor 4 (OCT- 4) was determined by real-time polymerase chain reaction (real time-PCR). nature renews itself tested by OCT- 4. 23R designed RIL- bioactivity evaluated by IL-17 and IL-10 expression of splenocytes of mice. cell differentiation has confirmed the isolation right-MSCs from lipoaspirate.
Restriction enzyme digestion and sequencing verified the successful cloning RIL- 23R in CD813A-1 lentiviral vectors. Green fluorescent protein (GFP) positive transduction level is up to 90%, and real-time PCR showed expression levels of RIL-23R. October-4 has the same expression pattern nontransduced-MSCs have and have-MSCs transduced / RIL-23R which showed that lentiviral vectors does not affect the characteristics of own-MSCs. Downregulation of IL-17, and upregulation of IL-10 showed activity right-MSCs have engineered.
The results show that transduced own-MSCs / RIL- 23R, express IL-23 decoy receptors, may provide a useful approach to basic research on cell-based gene therapy for autoimmune disorders.
The Human IL-23 Decoy Receptor Inhibits T-Cells Producing IL-17 by Genetically Engineered Mesenchymal Stem Cells.
Bioluminescent imaging exogenous Stem Cells in Intact Guinea Pig Cochlear.
Stem cell-based therapy can be used to replace damaged or missing neurons in the cochlear nerve of a patient suffering from sensorineural hearing loss severe-to-profound. In order to achieve functional recovery in future clinical trials, knowledge about the survival of transplanted cells and their differentiation into functional neurons is a prerequisite. It calls for non-invasive in vivo visualization of cell and long-term monitoring of life and their fate after transplantation cochlea. We have investigated if the molecular optical imaging allows visualization of cells within the cochlea exogenous intact cadaver heads guinea pigs.
Transduced (stem) cells stably co-expressing fluorescent (copGFP) and bioluminescent (Luc2) reporter molecules, which are injected into the internal auditory meatus or directly to the cochlea through the round window. After the injection of cells into the internal auditory meatus, glowing light signal observed in the cavum conchae of the ear, indicating that the light generated by Luc2 passing through the tympanic membrane and external auditory meatus.
pCDH-CMV-MCS-EF1-RFP cDNA Cloning and Expression Vector
Similar results were obtained after injection of the cells through the round window membrane, directly into the scala tympani canal or in Rosenthal in the modiolus of the cochlea basal turn. Imaging of the auditory bulla shows that bioluminescent signal passing through the tympanic membrane and the cracks in the wall of the bulla bone. After opening the auditory bulla, bioluminescent signal was coming from a round window. This is the first study showing that bioluminescence imaging allows visualization of luciferase-expressing cells injected into the intact guinea pig cochlea.
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