The microwell-mesh: A novel device and protocol for the high throughput manufacturing of cartilage microtissues

Futrega, Kathryn, Palmer, James S., Kinney, Mackenzie, Lott, William B., Ungrin, Mark D., Zandstra, Peter W., & Doran, Michael R. (2015) The microwell-mesh: A novel device and protocol for the high throughput manufacturing of cartilage microtissues. Biomaterials, 62, pp. 1-12.

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Abstract

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.

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8 citations in Web of Science®

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ID Code: 85865
Item Type: Journal Article
Refereed: Yes
Keywords: Differentiation, Stem cell, Cartilage, Mesenchymal stromal cell, Chondrocyte, 3D culture, Microtissue, Microwell
DOI: 10.1016/j.biomaterials.2015.05.013
ISSN: 1878-5905
Divisions: Current > Schools > School of Biomedical Sciences
Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2015 The Author(s)
Deposited On: 20 Jul 2015 23:57
Last Modified: 21 Jul 2015 21:56

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