International Journal of Stem Cells : eISSN 2005-5447

Fig. 2.

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Fig. 2. Type I collagen plays a crucial role in AM-1 tumoroid progression. (a) Brightfield image showing that round-shaped AM-1 tumoroids were observed in the Matrigel culture group. (b) Hematoxylin and eosin staining of a well-organized tumoroid structure, comprised of a heterogeneous cell population. (c) Col I was negatively expressed, and intense expression of Col IV was observed surrounding the edge of Matrigel-cultured tumoroids. (d) The terminal differentiation marker CK10 was strongly expressed in the tumoroid core, and p63 was intensely expressed in the basal cell layer. (e) MMP-9 showed a moderate expression in the basal cell layer. (f) E-cadherin was intensely expressed in the suprabasal layer and moderately expressed in the basal cell layer (indicated by a white dotted line). Vimentin was negatively expressed in the Matrigel culture group. (g) Net-like structures were observed in collagen gel-cultured (4 mg/ml) AM-1 tumoroids. (h) Hematoxylin and eosin staining of the invading strands from collagen gel-cultured tumoroids. (i) Immunohistochemistry staining of Col I and Col IV. Arrowheads indicate the rearranged fibrillar collagen fiber by adjacent AM-1 tumoroids, and Col IV was negatively expressed. (j) CK10 showed negative expression, and p63 showed a moderate expression. (k) Intense MMP-9 expression was observed in the majority of collagen-cultured AM-1 tumoroids. (l) E-cadherin showed a moderate expression, and vimentin was intensely expressed in the leading region of the invading tumor strand as indicated by an arrowhead. Nuclei were counterstained with TO-PRO-3 (TP-3). (m) Representative images of AM-1 cell cultured in Matrigel or varying concentrations of type I collagen gel (1, 2, and 4 mg/ml) obtained from 10 h of live-cell imaging. AM-1 cells were indicated with CMFDA dye (green). (n) Trajectory plots depicted the movement path of individual AM-1 cells (n=5). Through transformation, all trajectories started at the origin (x=0, y=0) in order to visualize the movements of all cells relative to each other. (o) Quantitative analysis of accumulation distance (μm) of Matrigel or collagen gel-cultured AM-1 cell. Scale bars: 100 μm. *p<0.05, ***p<0.001, and ****p<0.0001.
International Journal of Stem Cells 2022;15:415-21 https://doi.org/10.15283/ijsc22132
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