Stem cell-based repair auditory neurons may represent an attractive therapeutic option to restore sensorineural hearing loss. stem cells of hair-follicle-bulge-derived (HFBSCs) are promising candidates for this type of therapy, because they (1) have the nature of migration, allowing migration after transplantation, (2) can differentiate into sensory neurons and glial cells, and (3) can easily be harvested in relatively high amounts. However, HFBSCs never used for this purpose.
We hypothesize that HFBSCs can be used to repair the auditory nerve cell-based and we have researched their migration and incorporation into the cochlear modiolus explant and subsequent differentiation of them. Modiolus explants derived from adult wild type mice were cultured in the presence of HFBSCs EF1α-copGFP-transduced, constitutively expressing green fluorescent protein copepods (copGFP). Also, explants modiolus without hair cells were co-cultured with DCX-copGFP HFBSCs-transduced, indicating copGFP on doublecortin expression during neuronal differentiation.
HFBSC speed migration towards modiolus explants is calculated, and after two weeks, co-cultures were fixed and processed for immunohistochemical staining. EF1α-copGFP HFBSC rapid migration speed: 80.5 ± 6.1 m / h. After arriving in explants, cells form a pattern fasciculus and change their phenotype into neural cell types Atoh1-positive. DCX-copGFP HFBSCs green-fluorescent after integration into the explants, justify the differentiation of neurons from cells. These results indicate that HFBSC derived progenitor neuronal migration and can integrate into the cochlear modiolus explants, while adapting their phenotypes depending on the micro environment. Thus, HFBSCs demonstrate the potential to be employed in cell-based therapies to repair the auditory nerve.
The immunomodulatory and self-renewable human features adipose-derived mesenchymal stem cells (HAD-MSCs) mark their importance in regenerative medicine. Interleukin (IL) -23 as proinflammatory cytokines Hitting the regulatory T cells and promote the response of T helper 17 and T helper 1 cells. This initiation pathways of inflammation and immunosuppression in several autoimmune diseases.
Neuronal differentiation of hair-follicle-bulge-derived stem cells co-cultured with mouse cochlear modiolus explants.
[Construction of eukaryotic expression vector fibroblast activation protein and the formation of stable over-expression in squamous cell carcinoma of the mouth].
OBJECTIVE This study aimed to construct vectors fibroblast activation protein (FAP) over-expression lentivinus to investigate transfection in cell lines and build a stable FAP SCC9 over-expression of oral squamous cell lines.
METHOD The cDNA of the FAP gene from squamous cell carcinoma of the oral tissues (OSCC) was amplified by polymerase chain reaction (PCR) and subkloning into eukaryotic expression vector pCDH-CMV-MCS-EF1-copGFP. Recombinant plasmids were lined up and then transfected into cell lines SCC9. Further, cell lines which over-expressed FAP SCC9 stable founded by fluorescence activated cell sorting (FACS). Expression of green fluorescent protein (GFP) was detected with a fluorescence microscope, and over-expression of FAP identified by real-time PCR and Western blot.
RESULTS FAP gene amplified by PCR and then cloned into a vector, whose sequence is identical to that in GenBank. GFP was expressed in transfected cells. Furthermore, FAP over-expression in transfected cells was detected by real-time PCR and Western blot.
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: This product includes the reaction buffer, DNA, E. coli gyrase, ATP and the phosphate detection reagent for 100 assays of DNA supercoiling reactions in a 384-well assay format.
P. aeruginosa Gyrase ATPase assay Kit Plus (enzyme included)
Description: This product includes the reaction buffer, DNA, P. aeruginosa gyrase, ATP and the phosphate detection reagent for 100 assays of DNA supercoiling reactions in a 384-well assay format.
S. aureus gyrase ATPase assay Kit Plus (enzyme included)
Description: This product includes the reaction buffer, DNA, S. aureus gyrase, ATP and the phosphate detection reagent for 100 assays of DNA supercoiling reactions in a 384-well assay format.
CONCLUSION Recombinant eukaryotic expression vector pCDH-FAP successfully constructed. These results provide a basis for further study on the function of FAP in vitro.
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