科研之友
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摘要
Three-dimensional (3D) tissue models have significantly improved our understanding of structure/function relationships and promise to lead to new advances in regenerative medicine. However, despite the expanding diversity of 3D tissue fabrication methods, approaches for functional assessment have been relatively limited. Here, we describe the fabrication of microtissue (mu-tissue) suspensions and their quantitative evaluation with techniques capable of analyzing large sample numbers and performing multiplexed parallel analysis. We applied this platform to 3D mu-tissues representing multiple stages of liver development and disease including: embryonic stem cells, bipotential hepatic progenitors, mature hepatocytes, and hepatoma cells photoencapsulated in polyethylene glycol hydrogels. Multiparametric mu-tissue cytometry enabled quantitation of fluorescent reporter expression within populations of intact m-tissues (n >= 10(2)-10(3)) and sorting-based enrichment of subsets for subsequent studies. Further, 3D mu-tissues could be implanted in vivo, respond to systemic stimuli, retrieved and quantitatively assessed. In order to facilitate multiplexed 'pooled' experimentation, fluorescent labeling strategies were developed and utilized to investigate the impact of mu-tissue composition and exposure to soluble factors. In particular, examination of drug/gene interactions on collections of 3D hepatoma mu-tissues indicated synergistic influence of doxorubicin and siRNA knockdown of the anti-apoptotic gene BCL-XL. Collectively, these studies highlight the broad utility of mu-tissue suspensions as an enabling approach for high n, populational analysis of 3D tissue biology in vitro and in vivo.
- 出版日期2010
