Do you have suggested cell seeding protocols for different cell types?

As with any cultureware, optimal seeding density on Curi Bio Cytostretcher Chambers can vary according to cell type and the purpose of the experiment. However, we recommend researchers use the same cell density that they would use for conventional cultureware. For specific applications, please contact us at support@curibio.com.

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CytostretcherArianna Kieser
Is there a suggested cell seeding density for iPSCs?

For most commercially available cell lines, the cell seeding density used on conventional cultureware works with Curi Bio Cytostretcher Chambers as well. If you are using your own cell line, our support team can provide you with suggestions on how to design seeding density optimization experiments.

If you require additional support, please contact support@curibio.com so that our applications team can provide you with the relevant suggested protocol.

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CytostretcherArianna Kieser
Can I grow cells directly on Curi Bio Cytostretcher Chambers or do I need to do ECM protein coating?

We recommend coating the surface with the extracellular matrix protein you typically use in your cell culture workflow (collagen, fibronectin, gelatin, Matrigel, etc.). We also have a suggested concentration in our manual for protein coating of our substrates using some of these proteins. Before starting your experiments, make sure to check our manuals where you can find the details of the protocols for each protein coating.

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CytostretcherArianna Kieser
Cytostretcher Chambers come in flat or nanopatterned topographies. Which one should I use for my application?

Cytostretcher Chambers come in: (1) unpatterned "flat" and (2) NanoSurface topography. Both formats have a polymer substrate that is softer than conventional glass or plastic cultureware. As a result, the softer substrates are more representative of the mechanical properties of the tissue microenvironment. The NanoSurface substrate possesses submicron grooves with a feature size that is comparable to the collagen microenvironment in the body. As a result, the pattern is also optimized to be in the size of the focal adhesion of cells and facilitate the alignment of adherent cells. Our customers usually use flat substrates either for their improved mechanical properties (softer substrate) or as a control sample for NanoSurface topography to show the differences in the functionality of aligned and non-aligned tissue.

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CytostretcherArianna Kieser
Do I need a plasma machine to effectively adhere cells to the surface of Cytostretcher chambers?

Plasma treatment is one of the more effective methods for adequate surface modification. If you do not have access to a plasma machine in your lab, you can also use PDL coating. Please refer to our manual or contact our support team for details of each protocol.

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What protocols do you have for the use of the dishes. In the literature, several different seeding densities were used. What do you recommend?

As with any cultureware, optimal seeding density on ANFS can vary according to cell type and the purpose of the experiment. However, we recommend researchers use exactly the same cell density that they use for normal 35mm dishes, 6-well dishes and so on. Protocol information is provided on the ANFS product pages.

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Is there any type of pre-wash that must be performed prior to seeding the dishes?

No, ANFS is already gamma sterilized and plasma treated to promoted cellular adhesion. However, researchers may choose to deposit additional surface treatments depending on their exact cell type. Fibronectin, collagen, and PLL are all common surface treatment choices.

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