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Engineering human cell spheroids to model embryonic tissue fusion in vitro.
Citation:
Belair, D., Cynthiaj Wolf, C. Wood, H. Ren, R. Grindstaff, W. Padgett, A. Swank, D. MacMillan, A. Fisher, W. Winnik, AND B. Abbott. Engineering human cell spheroids to model embryonic tissue fusion in vitro. PLOS ONE . Public Library of Science, San Francisco, CA, 12(9):e0184155, (2017).
Impact/Purpose:
The purpose of this product was to establish, describe, and validate a method for studying the fusion of engineered tissues in vitro that is reminiscent of palate fusion. The data generated for the product suggest key cellular events and pathways needed to generate and validate an adverse outcome pathway for cleft palate. The method for assessing fusion behavior in vitro will enable future chemical testing to predict and prioritize which chemicals would likely interfere with palate fusion during development. This product demonstrates some validation of the in vitro model and identified that perturbation of either epidermal growth factor signaling or fibroblast growth factor signaling interferes with in vitro fusion and would be candidate pathways for identifying a set of chemicals from ToxCast that would likely interfere with fusion in vitro and in vivo.
Description:
Epithelial-mesenchymal interactions drive embryonic fusion events during development and upon perturbation can result in birth defects. Cleft palate and neural tube defects can result from genetic defects or environmental exposures during development, yet very little is known about the effect of chemical exposures on fusion defects in humans because of the lack of relevant and robust human in vitro assays of developmental fusion behavior. Given the etiology and prevalence of cleft palate and the relatively simple architecture and composition of the embryonic palate, we sought to develop a three-dimensional culture system that could be used to study fusion behavior in vitro using human cells. We engineered human Wharton’s Jelly stromal cell (HWJSC) spheroids of defined size and established that 7 days of culture in osteogenesis differentiation medium was sufficient to promote an osteogenic phenotype consistent with embryonic palatal mesenchyme. HWJSC spheroids supported the attachment of human epidermal keratinocyte progenitor cells on the outer spheroid surface likely through deposition of collagens I and IV, fibronectin, and laminin, and co-cultured spheroids exhibited fusion behavior that was dependent on epidermal growth factor signaling and fibroblast growth factor signaling in agreement with palate fusion literature. The method described here may broadly apply to the generation of three-dimensional epithelial-mesenchymal co-cultures to study developmental fusion events in a format that is amenable to predictive toxicology applications.
