Institute of Health and Biomedical Innovation, Queensland University of Technology at The Translational Research Institute, Brisbane, QLD, Australia.
Institute of Health and Biomedical Innovation, Queensland University of Technology at The Translational Research Institute, Brisbane, QLD, Australia; School of Chemistry, Physics and Mechanical Engineering, Science and Engineering Faculty, Queensland University of Technology Brisbane, QLD, Australia.
Biomaterials. 2015 Sep;62:1-12. doi: 10.1016/j.biomaterials.2015.05.013. Epub 2015 May 20.
Microwell platforms are frequently described for the efficient and uniform manufacture of 3-dimensional (3D) multicellular microtissues. Multiple partial or complete medium exchanges can displace microtissues from discrete microwells, and this can result in either the loss of microtissues from culture, or microtissue amalgamation when displaced microtissues fall into common microwells. Herein we describe the first microwell platform that incorporates a mesh to retain microtissues within discrete microwells; the microwell-mesh. We show that bonding a nylon mesh with an appropriate pore size over the microwell openings allows single cells to pass through the mesh into the microwells during the seeding process, but subsequently retains assembled microtissues within discrete microwells. To demonstrate the utility of this platform, we used the microwell-mesh to manufacture hundreds of cartilage microtissues, each formed from 5 × 10(3) bone marrow-derived mesenchymal stem/stromal cells (MSC). The microwell-mesh enabled reliable microtissue retention over 21-day cultures that included multiple full medium exchanges. Cartilage-like matrix formation was more rapid and homogeneous in microtissues than in conventional large diameter control cartilage pellets formed from 2 × 10(5) MSC each. The microwell-mesh platform offers an elegant mechanism to retain microtissues in microwells, and we believe that this improvement will make this platform useful in 3D culture protocols that require multiple medium exchanges, such as those that mimic specific developmental processes or complex sequential drug exposures.
微井平台常用于高效且均匀地制造 3 维（3D）多细胞微组织。多次部分或完全的培养基交换可以将微组织从离散的微井中置换出来，这可能导致微组织从培养中丢失，或者当被置换的微组织落入共同的微井中时发生微组织融合。本文中我们描述了第一个包含网格以保留离散微井内微组织的微井平台；微井-网格。我们展示了在微井开口上结合具有适当孔径的尼龙网格可以允许单细胞在接种过程中穿过网格进入微井，但随后保留组装好的微组织在离散的微井内。为了演示该平台的实用性，我们使用微井-网格制造了数百个软骨微组织，每个微组织由 5×10(3)个骨髓来源的间充质干细胞/基质细胞（MSC）形成。微井-网格在包括多次完全培养基交换的 21 天培养过程中可靠地保留了微组织。与由每个 2×10(5)MSC 形成的传统大直径对照软骨球相比，微组织中的软骨样基质形成更快且更均匀。微井-网格平台提供了一种将微组织保留在微井中的优雅机制，我们相信这种改进将使该平台在需要多次培养基交换的 3D 培养方案中非常有用，例如模拟特定发育过程或复杂顺序药物暴露的方案。
