Reproducible, ultra high-throughput formation of multicellular organization from single cell suspension-derived human embryonic stem cell aggregates.
Reproducible, ultra high-throughput formation of multicellular organization from single cell suspension-derived human embryonic stem cell aggregates.
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DOI：
10.1371/journal.pone.0001565
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发表时间：
2008-02-13
期刊：
影响因子：
3.7
通讯作者：
Zandstra PW
中科院分区：
文献类型：
--
作者：
Ungrin MD;Joshi C;Nica A;Bauwens C;Zandstra PW
人类胚胎干细胞（hESC）应该使新的见解，早期人类发育，并提供再生医学细胞的可再生来源。然而，由于通常用于揭示hESC发育潜力的三维hESC聚集体[胚状体（hEB）]是异质的并且表现出无序的分化，因此hESC技术开发的进展受到阻碍。使用离心强制聚集策略结合一种新的离心提取方法作为基础，我们证明了hESC输入组合物和诱导环境可以被操纵以形成大量的明确的聚集体，其表现出多谱系分化和从单细胞悬浮液中显著改善的自组织。这些聚集体表现出协调的双域结构，包括胚外内胚层和外胚层样组织的连续区域。使用基于硅晶片的微制造技术来产生允许每平方厘米产生数百至数千个hEB的表面。早期人类胚胎发生的机制知之甚少。我们报告了一种用于产生空间和时间同步的hEB的超高通量（UHTP）方法。以这种方式产生的聚集体表现出植入周围组织水平形态发生的方面。这些结果应该推进对早期人类发育过程的基础研究，使高通量筛选策略能够识别指定hESC衍生细胞和组织的条件，并加速hESC衍生细胞的临床前评估。
Human embryonic stem cells (hESC) should enable novel insights into early human development and provide a renewable source of cells for regenerative medicine. However, because the three-dimensional hESC aggregates [embryoid bodies (hEB)] typically employed to reveal hESC developmental potential are heterogeneous and exhibit disorganized differentiation, progress in hESC technology development has been hindered. Using a centrifugal forced-aggregation strategy in combination with a novel centrifugal-extraction approach as a foundation, we demonstrated that hESC input composition and inductive environment could be manipulated to form large numbers of well-defined aggregates exhibiting multi-lineage differentiation and substantially improved self-organization from single-cell suspensions. These aggregates exhibited coordinated bi-domain structures including contiguous regions of extraembryonic endoderm- and epiblast-like tissue. A silicon wafer-based microfabrication technology was used to generate surfaces that permit the production of hundreds to thousands of hEB per cm2. The mechanisms of early human embryogenesis are poorly understood. We report an ultra high throughput (UHTP) approach for generating spatially and temporally synchronised hEB. Aggregates generated in this manner exhibited aspects of peri-implantation tissue-level morphogenesis. These results should advance fundamental studies into early human developmental processes, enable high-throughput screening strategies to identify conditions that specify hESC-derived cells and tissues, and accelerate the pre-clinical evaluation of hESC-derived cells.
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影响因子：
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