Sheridan Julie M, Taoudi Samir, Medvinsky Alexander, Blackburn C Clare
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, University of Edinburgh, United Kingdom.
Genesis. 2009 May;47(5):346-51. doi: 10.1002/dvg.20505.
Cellular reaggregation methods have been used to generate in vitro organotypic cultures as a means to elucidate the cellular and molecular requirements of organogenesis. However, reproducibility from experiment to experiment has remained problematic and furthermore, current protocols do not support reaggregation of many important tissues. Here, using the thymus as a model organ, we present a novel reaggregation method termed "compaction reaggregation" that offers improved kinetics of reaggregation and greatly improved efficiency. Using compaction reaggregation we have been able to reaggregate the aorta-gonad- mesonephros region, a tissue that previously proved refractory to commonly used reaggregation methods, enabling the study of hematopoietic stem cell emergence and expansion. Additionally, compaction reaggregation permits the juxtaposition of different cell layers within the aggregated structure thus providing the means to study inductive interactions between different cell populations in vitro.
细胞重聚集方法已被用于生成体外器官型培养物，以此作为阐明器官发生的细胞和分子需求的一种手段。然而，实验之间的可重复性一直存在问题，此外，目前的方案并不支持许多重要组织的重聚集。在这里，我们以胸腺作为模型器官，提出了一种名为“压实重聚集”的新型重聚集方法，该方法具有更快的重聚集动力学且效率大大提高。使用压实重聚集，我们能够使主动脉-性腺-中肾区域重聚集，该组织以前被证明对常用的重聚集方法具有抗性，从而能够研究造血干细胞的出现和扩增。此外，压实重聚集允许在聚集结构内并列不同的细胞层，从而提供了在体外研究不同细胞群体之间诱导相互作用的手段。
