Embryonic stem (ES) cells provide a potentially unlimited source of specialized cells for regenerative medicine. The ease of inducing stable genetic modifications in ES cells allows for in vitro manipulations to enhance differentiation into specific cell types and to optimize in vivo function of differentiated progeny in animal models of disease. We have generated mouse ES cells that constitutively express Bcl-XL, an antiapoptotic protein of Bcl-2 family. In vitro differentiation of Bcl-XL overexpressing ES (Bcl-ES) cells resulted in higher expression of genes related to midbrain dopamine (DA) neuron development and increased the number of ES-derived neurons expressing midbrain DA markers compared with differentiation of wild-type ES cells. Moreover, DA neurons derived from Bcl-ES cells were less susceptible to 1-methyl-4-phenylpyridium, a neurotoxin for DA neurons. On transplantation into parkinsonian rats, the Bcl-ES-derived DA neurons exhibited more extensive fiber outgrowth and led to a more pronounced reversal of behavioral symptoms than wild-type ES-derived DA neurons. These data suggest a role for Bcl-XL during in vitro midbrain DA neuron differentiation and provide an improved system for cell transplantation in a preclinical animal model of Parkinson's disease.
Background
SARS‐CoV‐2 pandemic is currently ongoing, meanwhile vaccinations are rapidly underway in some countries. The quantitative immunoassays detecting antibodies against spike antigen of SARS‐CoV‐2 have been developed based on the findings that they have a better correlation with the neutralizing antibody.
Methods
The performances of the Abbott Architect SARS‐CoV‐2 IgG II Quant, DiaSorin LIAISON SARS‐CoV‐2 TrimericS IgG, and Roche Elecsys anti‐SARS‐CoV‐2 S were evaluated on 173 sera from 126 SARS‐CoV‐2 patients and 151 pre‐pandemic sera. Their correlations with GenScript cPass SARS‐CoV‐2 Neutralization Antibody Detection Kit were also analyzed on 173 sera from 126 SARS‐CoV‐2 patients.
Results
Architect SARS‐CoV‐2 IgG II Quant and Elecsys anti‐SARS‐CoV‐2 S showed the highest overall sensitivity (96.0%), followed by LIAISON SARS‐CoV‐2 TrimericS IgG (93.6%). The specificities of Elecsys anti‐SARS‐CoV‐2 S and LIAISON SARS‐CoV‐2 TrimericS IgG were 100.0%, followed by Architect SARS‐CoV‐2 IgG II Quant (99.3%). Regarding the correlation with cPass neutralization antibody assay, LIAISON SARS‐CoV‐2 TrimericS IgG showed the best correlation (Spearman rho = 0.88), followed by Architect SARS‐CoV‐2 IgG II Quant and Elecsys anti‐SARS‐CoV‐2 S (all rho = 0.87).
Conclusions
The three automated quantitative immunoassays showed good diagnostic performance and strong correlations with neutralization antibodies. These assays will be useful in diagnostic assistance, evaluating the response to vaccination, and the assessment of herd immunity in the future.
Wharton's jelly is not only one of the most promising tissue sources for mesenchymal stem cells (MSCs) but also a source of natural growth factors. To prove that we can get both natural growth factors and MSCs from Wharton's jelly, we compared cellular characteristics and the level of basic fibroblast growth factor (bFGF) from samples using the explant culture method to those derived from the traditional enzymatic culture method. The levels of bFGF were 27.0 ± 11.7 ng/g on day 3, 15.6 ± 11.1 ng/g on day 6, and decreased to 2.6 ± 1.2 ng/g on day 14. The total amount of bFGF released was 55.0 ± 25.6 ng/g on explant culture. Compared with the traditional enzymatic digestion method, the explant culture method showed a tendency to release higher levels of bFGF in supernatant media for the first week of culture, and the higher cellular yield at passage 0 (4.89 ± 3.2 × 105/g versus 1.75 ± 2.2 × 105/g, P = 0.01). In addition, the genes related to mitosis were upregulated in the explant-derived MSCs.
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