Journal: Journal of Medical Virology

Article Title: SARS‐CoV‐2 pseudovirus infectivity and expression of viral entry‐related factors ACE2, TMPRSS2, Kim‐1, and NRP‐1 in human cells from the respiratory, urinary, digestive, reproductive, and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Cell lines used in the present study

Article Snippet: BC‐3 , Primary effusion lymphoma , ATCC CRL‐2277 , RPMI‐1640 complete medium.

Techniques: Cell Culture

Journal: Scientific reports

Article Title: Co-aggregation of MSC/chondrocyte in a dynamic 3D culture elevates the therapeutic effect of secreted extracellular vesicles on osteoarthritis in a rat model.

doi: 10.1038/s41598-022-22592-4

Figure Lengend Snippet: Figure 2. qRT-PCR analysis of chondrogenic genes. (A) Histogram displays the qRT-PCR analysis of collagen type II (Col II), Sox9, aggrecan (Acan), and Col X in a cell micromass culture in the existence of extracellular vesicles (EVs) derived from chondrocyte aggregates (Cho-ag-EV) and MSC aggregates (MSC-ag-EV) and Co-aggregates (Co-ag-EV) in chondrogenic medium with ( +) and without (−) TGFß1 after 21 days in vitro histological analysis. (B, a) Statistical analysis of the comparison between micromass diameters of all experimental groups and the control that demonstrated the Co-ag-EV + and MSC-ag-EV + groups show bigger size and differentiation phenotype of MSCs to chondrocytes than the control group (received chondrogenic medium contain TGFß1) and Cho-ag-EV group. (B, b) Toluidine blue was performed for (TB) of mesenchymal stem cells (MSCs) micromass sections that differentiated into chondrocytes in the presence of extracellular vesicles (EVs) harvested from chondrocyte aggregates (Cho-ag-EV) and MSC aggregates (MSC-ag-EV) and Co-aggregates (Co-ag-EV) in chondrogenic medium with ( +) and without (−) TGFß1 after 21 days. (B, c) Statistical analysis of TB staining based on the percent of positive staining area. TB staining does not display a statistically significant difference between the groups. (B, d) safranin O (SO) staining was performed for all micromass sections. (B, e) Statistical analysis of SO staining based on the percent of positive staining area. SO staining does not display a statistically significant difference between the groups. The results are expressed as the mean ± SEM. *Significant difference compared with the control group (*p < 0.05; **p < 0.01).

Article Snippet: To assess the chondrogenic proteins expression level in the extracted EVs, we also executed a wet western blot analysis with the primary antibodies of COL II (COL2A1, 250,484, Abbiotec, USA), aggrecan (ACAN, NB600-504, Novus Biologicals, USA), and COL X (COL10A, ab58632, Abcam, UK).

Techniques: Quantitative RT-PCR, Derivative Assay, In Vitro, Comparison, Control, Staining

Journal: Cells

Article Title: Interaction Between Human Skeletal and Mesenchymal Stem Cells Under Physioxia Enhances Cartilage Organoid Formation: A Phenotypic, Molecular, and Functional Characterization

doi: 10.3390/cells14181423

Figure Lengend Snippet: Extracellular matrix (ECM) protein production in cartilage organoids: ( A ) Immunohistochemical detection of aggrecan, type II collagen, proteoglycan-4, type I collagen, and type X collagen in SSC-derived organoids cultured without (–IM) or with (+IM) chondrogenic induction medium. Images acquired with an Olympus microscope at 10× magnification. ( B ) Immunohistochemical detection of the same ECM proteins in MSC-derived organoids under the same conditions. ( C ) Immunohistochemical detection of ECM proteins in organoids derived from SSC–MSC co-cultures, with or without chondrogenic induction medium. Images acquired with an Olympus microscope at 10× magnification. ( D – F ) Semi-quantitative scoring of ECM protein expression (aggrecan, type II collagen, proteoglycan-4, type I collagen, and type X collagen) in SSC-derived organoids ( D ), MSC-derived organoids ( E ), and SSC–MSC co-culture organoids ( F ), cultured with or without chondrogenic induction medium. ( G ) Comparative scoring of aggrecan, type II collagen, and proteoglycan-4 expression between cartilage organoids and the NHAC-kn chondrocyte line. ( H ) Comparative scoring of type I and type X collagen expression between cartilage organoids and the NHAC-kn chondrocyte line. The statistical difference is generated by Group A ( G , H ). Statistical significance was determined using the Kruskal–Wallis test followed by multiple comparisons; * p < 0.05; *** p < 0.001; **** p < 0.0001. ns: non-significant.

Article Snippet: For immunohistochemistry (IHC), the Autostainer Link 48 automated system was used with the following antibodies: aggrecan (Clone BC-3, Novus ® , Novus Biologicals, Centennial, CO, USA), collagen I alpha 1 (Clone COL-1, Novus ® , Novus Biologicals, Centennial, CO, USA), collagen II (Clone 5B2.5, Novus ® , Novus Biologicals, Centennial, CO, USA), collagen X alpha 1 (Clone SR3302, Novus ® , Novus Biologicals, Centennial, CO, USA), and PRG4 (NBP1-19048, Novus ® , Novus Biologicals, Centennial, CO, USA).

Techniques: Immunohistochemical staining, Derivative Assay, Cell Culture, Microscopy, Expressing, Co-Culture Assay, Generated