Скачать или смотреть Preparing BD Matrigel™-coated plate for STEMium® use

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Still image of BD Matrigel bottle next to our DMEM F12 In this video, we will demonstrate how to prepare BD Matrigel-coated plate for feeder-free culturing of human pluripotent stem cells in STEMium. It is important that users become thoroughly familiar with the entire method before proceeding

List of reagents The following reagents are required: DMEM F12, complete STEMium, an aliquot of BD Matrigel, a six well plate, pipettors, pipette tips and serological pipettes, laminar flow hood and a 37 degree celsius 5% carbon dioxide incubator.

Diluting Matrigel Video To begin, follow the maunufacturer's instruction by thawing BD Matrigel overnight at 4 degrees celsius. Next, transfer cold DMEM F12 to a prechilled conical tube. An aliquot of thawed BD Matrigel is then added to the DMEM F12.

Coating Plate Video Shown here is the prepared working solution of BD Matrigel. To coat plate, pipette 1mL of the BD Matrigel working solution per well and distribute evenly. Incubate plate at 37 degrees celsius for 30 minutes.

After incubation, remove the BD Matrigel coated plate and aspirate the remaining solution from well while avoiding contact with the substrate.

Next, add 2mLs of complete STEMium and the plate is ready for use. Plate can be left in 37 degrees celsius incubator during cell passaging. For instructions on how to passage human pluripotent stem cells under feeder-free conditions, please see our online video library.


Helpful Suggestions
We recommend the following suggestions:
It is important to not warm BD Matrigel solution to prevent premature gelling, which may decrease its properties
For optimal culture, ensure that the surface is evenly coated with BD Matrigel substrate.
Prepare BD Matrigel-coated plate immediately prior to use, preferably an hour before.


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