Hi, I wanted to perform tube formation assay using HUVEC cells. I want to use Corning Matrigel (High protein) for the assay. As per the manufacturer's instructions, I have thawed the Matrigel at 4 degree C overnight and it looked almost liquid very next morning. I also followed the instructions to pre-cool pipette tips and tubes used for the assay. But while aliquoting in ice, the Matrigel became gel-like and was not even coming from the tip. I did not dilute the Matrigel with any media.
Any suggestions how to go ahead for aliquoting and subsequent coating of Matrigel into Ibidi micro angiogenesis plates. I have to use 10 micro L for coating but the gel-like Matrigel is making it difficult. Thanks in advance.
Hi Gejo,
To us it sounds like you did everything right. However, it is possible that Matrigel could polymerize - even with all the precautions you took, - if the ambient temperature of the room is too high. What you could try is thawing the Matrigel on ice in the refrigerator, too prevent it from polymerizing at this step. Please take a look at the ibidi Application Note 19 (https://ibidi.com/img/cms/support/AN/AN19_Tube_Formation.pdf) where we describe how we apply Matrigel to the µ-Slide Angiogenesis. For any additional tips I would suggest you visit the FAQs from Corning, you might find something useful there.
Hi Tina.
Thank you for your suggestions. We had to work in room temperature without air condition in our laboratory, probably that could have been the reason. From last weekend, I could trouble shoot this issue to a satisfactory level following the above guidelines you mentioned. Hoping to have your support in this experiment if need arises.
That's great news. Please get in touch, if you have further questions: [email protected]
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