Coating Plates with Matrigel® for Pluripotent Stem Cell Culture
Culturing and maintaining human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs) with mTeSR™1 or TeSR™2 medium requires the use of tissue culture plates coated with a matrix to support cell attachment and expansion. Corning® Matrigel® hESC-qualified Matrix (Corning Catalog #354277) is recommended for coating tissue culture plates. The following tips provide recommendations for coating plates with Matrigel®.
The diluting instructions for Matrigel® hESC-qualified Matrix are provided in the Product Insert. The dilution factor is calculated for each lot of Matrigel®, based on the protein concentration. Please refer to the lot-specific Certificate of Analysis, which can be requested on the Corning® website.
Ensuring Even Coating
It is important that each tissue culture plate is completely and evenly coated with Matrigel®. Cells will not attach properly where there is insufficient Matrigel® coating. One way to avoid uneven coating is to let the coated plates sit in a biological safety cabinet at room temperature for 1 hour with the lid on (to prevent dehydration). If you are in a rush, plates can be coated for 30 minutes in a 37 °C incubator instead. Make sure that the surface of the safety cabinet or incubator shelf is level (use a bubble level for confirmation) and avoid stacking the plates on top of each other.
Storage of Matrigel® Coated Plates
Coated plates can be stored at 2-8 °C for up to one week (sealed with Parafilm™ to prevent dehydration) before use. Make sure that the fridge shelf is level as well. We do not recommend using plates that have areas of uneven Matrigel® coating or plates where the Matrigel® has evaporated.
For more information about culturing pluripotent stem cells in mTeSR™1, please refer to the Technical Manual: Maintenance of Human Pluripotent Stem Cells in mTeSR™1.
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