Automated selection and collection of pluripotent stem cell colonies using the CellCelector™

Haupt, Simone; Grützner, Jan; Rath, Barbara H.; Möhlig, Heike; Brüstle, Oliver

doi:10.1038/nmeth.f.252

Cited by 6 publications

(4 citation statements)

References 3 publications

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“…Here, we report an optimized process for the automated transfer of individual antibody‐secreting cells from dense arrays of subnanoliter microwell containers into conventional 96‐well microtiter plates. The approach uses a commercially available instrument designed for colony picking26 and a custom software module to translate data obtained from microengraving—a soft lithographic method for high‐throughput, single‐cell screening27, 28—into a list of positions from which to pick the desired cells. The locations of wells containing clones of interest were identified from the areas of captured antibodies detected on processed microarrays, and these data guided the automated retrieval of those cells.…”

Section: Introductionmentioning

confidence: 99%

Development and optimization of a process for automated recovery of single cells identified by microengraving

Choi

Ogunniyi

et al. 2010

Biotechnology Progress

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Microfabricated devices are useful tools for manipulating and interrogating large numbers of single cells in a rapid and cost-effective manner, but connecting these systems to the existing platforms used in routine high-throughput screening of libraries of cells remains challenging. Methods to sort individual cells of interest from custom microscale devices to standardized culture dishes in an efficient and automated manner without affecting the viability of the cells are critical. Combining a commercially available instrument for colony picking (CellCelector, AVISO GmbH) and a customized software module, we have established an optimized process for the automated retrieval of individual antibody-producing cells, secreting desirable antibodies, from dense arrays of subnanoliter containers. The selection of cells for retrieval is guided by data obtained from a high-throughput, single-cell screening method called microengraving. Using this system, 100 clones from a mixed population of two cell lines secreting different antibodies (12CA5 and HYB099-01) were sorted with 100% accuracy (50 clones of each) in approximately 2 h, and the cells retained viability.

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Section: Introductionmentioning

confidence: 99%

Development and optimization of a process for automated recovery of single cells identified by microengraving

Choi

Ogunniyi

et al. 2010

Biotechnology Progress

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“…The precision and biological integrity of the Cell X TM biopsy/pick/week operations represents a significant improved over previous devices which pick cells from 2D surfaces by scrapping them off of a surface (Haupt et al 2009;Zoldan et al 2010). The Cell X TM method of pulling cells off of the surface using sheer force, prevents crushing or smearing of cells, which may compromise cell yield, cell viability, and the composition of bioactive factor in the sample after replating.…”

Section: Resultsmentioning

confidence: 99%

Automated in-process characterization and selection of cell-clones for quality and efficient cell manufacturing

et al. 2020

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Delivery of safe, effective and reliable cellular therapies, whether based on mesenchymal stromal cells (MSCs) or induced pluripotent stem cells (iPSCs), demand standardization of cell culture protocols. There is a need to develop automation platform that enables the users to generate culture expanded human cell populations that improves the quality and reduces batch-to-batch variation with respect to biological potential. Cell X TM robot was designed to address these current challenges in the cell fabrication industry. It utilizes non-invasive large field of view quantitative image analysis to guide an automated process of targeted ''biopsy'' (cells or media), ''picking'' (selection) of desired cells or colonies, or ''weeding'' (removal) of undesired cells, thus providing an unprecedented ability to acquire quantitative measurement in a complex heterogeneous cell environment ''in process'' and then to act on those measurements to define highly reproducible methods for cell and colony ''management'' based on application specific critical quality attributes to improve the quality of the manufactured cell lines and cell products.

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“…In a special experiment, the ability of this simple approach to yield high-quality cell preparations was confirmed by a flow cytometry analysis of the selected colonies (Supplementary Figure S6). Yet it is conceivable that the throughput and reliability of the procedure could be further improved by using automated devices for colony imaging and collection (36). …”

Section: Resultsmentioning

confidence: 99%

Planar Arrangement of Eukaryotic Cells in Merged Hydrogels Combines the Advantages of 3-D and 2-D Cultures

2012

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We report an unordered 2-D array of eukaryotic cells completely embedded in a 3-D matrix. Every cell is located at the same distance from the gel surface, which ensures uniformity of growth conditions and ease of observation characteristic of a 2-D culture. Each cell is firmly immobilized, and each has a unique address in the array. The cells can be rapidly screened, individually monitored during extended time periods, and cultured with the formation of spheroid microcolonies characteristic of a 3-D culture. Individual microcolonies can be extracted from the gel and further propagated, thus enabling isolation of pure cell clones from rather dense cell populations and rapid drug-free generation of stable cell lines.

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Automated selection and collection of pluripotent stem cell colonies using the CellCelector™

Cited by 6 publications

References 3 publications

Development and optimization of a process for automated recovery of single cells identified by microengraving

Development and optimization of a process for automated recovery of single cells identified by microengraving

Automated in-process characterization and selection of cell-clones for quality and efficient cell manufacturing

Planar Arrangement of Eukaryotic Cells in Merged Hydrogels Combines the Advantages of 3-D and 2-D Cultures

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