Journal: Molecular Biology Reports

Article Title: Optimising the method to develop spheroids from MDA-MB-468 human triple negative breast cancer cells

doi: 10.1007/s11033-026-11451-4

Figure Lengend Snippet: 3D culture formation was assessed using agarose-coated 96-well plates under two oxygen conditions: normoxic (NOC) [A] and low oxygen conditions (LOC) [D]. Morphometric parameters, including solidity, sphericity, and perimeter, were quantified using AnaSP image analysis software. Results for spheroids formed under NOC are shown in panels [B] and [C], and under LOC in panels [E] and [F]. Images are representative of a single well that was followed longitudinally across the captured time period; Red arrows indicate cellular debris. Data are mean ± SEM from three independent biological replicates, with > 3 spheroids per condition. Statistical significance: p < 0.05 (*) and p < 0.01 (**) using one -way ANOVA. Images were captured using an inverted light microscope (Nikon Eclipse TE2000-U) at 4× magnification. 3D: three dimensional ; NOC: normal oxygen condition; LOC: Low oxygen condition; scale bar = 200 micron

Article Snippet: Uniform aggregates were then placed into at the with same aggregates formed at the same time points with around 10 minuts difference for each replica (i) agarose-coated 96-well plates (JetBiofil, Cat. No.CAP011096, China) coated with 50 μL 1% agarose/well [ ], or (ii) Ultra–low-attachment (ULA) 96-well plates (Corning ® , Cat. No. 4515, Corning, USA).

Techniques: Software, Light Microscopy

Journal: Molecular Biology Reports

Article Title: Optimising the method to develop spheroids from MDA-MB-468 human triple negative breast cancer cells

doi: 10.1007/s11033-026-11451-4

Figure Lengend Snippet: qPCR analysis of gene expression in 3D spheroids generated under normoxic and low-oxygen conditions. Relative mRNA levels of CD44 , HIF1A , NES , SNAl1 , TWIST1 and VEGFA were quantified by qPCR in MDA-MB-468 spheroids formed using [A - B] agarose-coated 96-well culture plate, [C- D] ULA-plate and [E , F] scaffold plate under NOC [ A , C , E] and LOC [B , D , F]. Expression was assessed on days 4, 5, and 6 post-seeding and compared to 2D (control), [G] Day-6 heat map comparing log₂ expression of CD44 , HIF1A , NES , SNAI1 , TWIST1 , and VEGFA across all conditions (agarose, ULA, scaffold; NOC and LOC). Colour scale: yellow = higher expression, purple = lower expression. Values are means of n = 3 biological replicates. Two-way or one-way ANOVA was applied, as appropriate. Statistical significance was set at p < 0.05 (*) and p < 0.01 (**), p < 0.001(***), p < 0.0001(****) D=Days; ULA-plate: ultra-low attachment 96well plate; NOC: normal oxygen condition; LOC: Low oxygen condition

Article Snippet: Uniform aggregates were then placed into at the with same aggregates formed at the same time points with around 10 minuts difference for each replica (i) agarose-coated 96-well plates (JetBiofil, Cat. No.CAP011096, China) coated with 50 μL 1% agarose/well [ ], or (ii) Ultra–low-attachment (ULA) 96-well plates (Corning ® , Cat. No. 4515, Corning, USA).

Techniques: Gene Expression, Generated, Expressing, Control