Abstract
Mesenchymal stem cell spheroids have been encapsulated in hydrogels for various applications because spheroids demonstrate higher cell activity compared to individual cells in suspension. However, there is limited information about the effect of distance between spheroids (inter-spheroid distance) on fusion or migration in hydrogel. In this study, we developed temperature-responsive hydrogels with surface microwell patterns to culture adipose-derived stem cell (ASC) spheroids and deliver them into Matrigel for investigation of the effect of inter-spheroid distance on spheroid behavior. The ASC spheroids were encapsulated successfully in Matrigel, denoted as sandwich-culture, with specific inter-spheroid distance ranging from 100 to 400 µm. Interestingly, ASCs migrated from the host spheroid and formed a bridge-like structure between spheroids, denoted as a cellular bridge, only when the inter-spheroid distance was 200 µm. Thus, we sandwich-cultured human umbilical vein endothelial cells (HUVECs) and ASC in co-cultured spheroids in Matrigel to create a homogeneous endothelial cell network in the hydrogel. The HUVECs sprouted through the ASC cellular bridge and directly interacted with the adjacent spheroid when inter-spheroid distance was 200 µm. Similar results were obtained from in vivo study. Thus, our study suggests the appropriate inter-spheroid distance for effective spheroid encapsulation in hydrogel.
Published Version
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