Figure 3.

In vitro tube formation analysis of tumour cell-derived vascular mimicry (VM). (A) The Angiogenesis Analyser plugin was used to discern the optimal starting cell density needed to establish VM meshwork in vitro. Different tube formation parameters were analysed, including the number of junctions, segments, meshes, total mesh area, total segment length, and total branching lengths of the tubular networks. A starting cell density of 60 × 10³ produced consistent mature looping patterns in Matrigel for almost all the included cell lines. However, overall, the analysis shows that both 40 and 60 × 10³ concentrations are sufficient to initiate VM structures in vitro. (B) The results were comparable in Myogel, with better tubes formed with a starting cell density of 60 × 10³. However, at such a higher density, HUVEC meshwork became more extensively interlaced in Myogel, which limited the analyser’s capacity to recognize smaller tubular areas as shown in the figure. Data from one representative experiment, presented as mean ± SD of three technical replicates, are shown.