Journal: BMC Research Notes

Article Title: Development of a high throughput drug screening assay to identify compounds that protect oligodendrocyte viability and differentiation under inflammatory conditions

doi: 10.1186/s13104-016-2219-8

Figure Lengend Snippet: IFNγ reduces OL viability and inhibits differentiation. A Expanded OPCs were plated in 96 well plates in DMEM differentiation media for 24 h and either treated for 1 h with 1.1 µM quetiapine (QTP) or vehicle (0.1 % DMSO), followed by 48 h of 200 U/ml INFγ insult. No Insult was vehicle treated with no IFNγ insult. OLs were immunostained for MBP ( green ) and nuclei stained with DAPI ( blue ). Bar 200 μM. B 24 h after plating, differentiating OLs were treated with increasing concentrations of INFγ for 48 h. alamarBlue® (AB) fluorescence was quantified to determine cell viability. C 24 h after plating, differentiating OLs were treated with increasing concentrations of QTP for 1 h followed by 48 h treatment with 200 U/ml IFNγ. Cell viability was measured by quantification of alamarBlue® fluorescence. Image quantification of anti-MBP immunostaining of the same cultures was used to measure OL differentiation

Article Snippet: Paraformaldehyde solution was purchased from Electron Microscopy Science (Hatfield, PA). alamarBlue® (AB) was purchase from AbD Serotech (Killington, UK).

Techniques: Staining, Fluorescence, Immunostaining

Journal: BMC Research Notes

Article Title: Development of a high throughput drug screening assay to identify compounds that protect oligodendrocyte viability and differentiation under inflammatory conditions

doi: 10.1186/s13104-016-2219-8

Figure Lengend Snippet: TNFα synergizes with IFNγ causing reduced OL survival and inhibiting differentiation. Expanded OPCs were plated in DMEM differentiation media for 24 h and then treated with various combinations cytokines for 48 h. A Differentiating OLs were treated with increasing concentrations of TNFα alone ( gray bars ) or increasing concentrations of TNFα + 100 U/ml INFγ ( black bars ) and alamarBlue® (AB) fluorescence was quantified to determine cell viability. B Cells were treated as in A with increasing concentrations of TNFα alone ( black bars ) or TNFα + 5, 10, or 20 U/ml INFγ ( light gray , dark gray , or white bars respectively) and AB fluorescence was quantified to determine cell viability. C Image quantification of anti-MBP immunostaining of the same cultures as in B was used to measure OL differentiation. D Comparison of MBP expression in untreated (No insult) or 10 U/ml INFγ + 1 ng/ml TNFα treated differentiating OLs. E Viability and OL differentiation raw values of no insult versus 10 U/ml INFγ + 1 ng/ml TNFα demonstrate the reproducibility of the combined cytokine toxicity. Coefficient of variation (CV) values for cell viability were 7.47 and 6.14 % and for OL differentiation were 11.37 and 19.16 % for No insult or TNFα+INFγ, respectively. CV values <20 % were considered in the acceptable range

Article Snippet: Paraformaldehyde solution was purchased from Electron Microscopy Science (Hatfield, PA). alamarBlue® (AB) was purchase from AbD Serotech (Killington, UK).

Techniques: Fluorescence, Immunostaining, Expressing

Journal: BMC Research Notes

Article Title: Development of a high throughput drug screening assay to identify compounds that protect oligodendrocyte viability and differentiation under inflammatory conditions

doi: 10.1186/s13104-016-2219-8

Figure Lengend Snippet: Flow scheme of the cytokine OL protection assay. A primary O4-positive OPCs were isolated for P6–P7 neonatal rat brains. A OPCs were expanded in flasks for 5–7 days in defined media supplemented with PDGF. B Expanded OPCs were plated in 96-well plates in defined media supplemented with T3, but without PDGF. C Test compounds were added following a 24-h incubation period followed 1 h later by addition of cytokine challenge (1 ng/ml TNFα+10 U/ml IFNγ). D After 48-h culture, alamarBlue® (AB) is added and AB fluorescence used to determine viability and metabolic activity of the cells. After selection of hit compounds (>50 % of the positive control, 1.1 µM QTP), fresh compound from a new source was obtained and the dose response relationship and EC 50 values of each compound were determined. Following AB read out of the dose response, cells were fixed, immunostained, and images quantified to determine OL differentiation (MBP expression)

Article Snippet: Paraformaldehyde solution was purchased from Electron Microscopy Science (Hatfield, PA). alamarBlue® (AB) was purchase from AbD Serotech (Killington, UK).

Techniques: Isolation, Incubation, Fluorescence, Activity Assay, Selection, Positive Control, Expressing

Journal: Scientific Reports

Article Title: Controlling osteoblast morphology and proliferation via surface micro-topographies of implant biomaterials

doi: 10.1038/s41598-020-69685-6

Figure Lengend Snippet: ( a ) AlamarBlue reduction and ( b ) DNA concentration of AO after 1 and 7 days of culture. Data show mean values ± SEM (n = 6 per group). Statistically significant differences (p < 0.05, Tukey’s HSD test) were marked with “a” if a Ce-TZP surface differed statistically significant from the corresponding Y-TZP surface, with “b” if Y-TZP + CaP, Y-TZPgr or Y-TZPsa differed from Y-TZP or if Ce-TZP + CaP, Ce-TZPgr or Ce-TZPsa differed from Ce-TZP, with “c” if Y-TZPgr or Y-TZPsa differed from Y-TZP + CaP or if Ce-TZPgr or Ce-TZPsa differed from Ce-TZP + CaP and with “d” if Y-TZPsa differed from Y-TZPgr or if Ce-TZPsa differed from Ce-TZPgr. *Statistically significant difference between day 1 and day 7.

Article Snippet: The metabolic activity of AO cultured on the test surfaces was analysed by the alamarBlue (AB) metabolic assay (MorphoSys AbD, Düsseldorf, Germany).

Techniques: Concentration Assay