Hello. I am trying to generate monolayers from cow enteroids. The protocol I am following involves 0.5uM EDTA and Trypsin-EDTA 0.05%, each enzyme is applied in two different steps. But for some reason the Matrigel is not disrrupted and remains within the pellet I obtained after centrifugation. Does anyone has experience using Corning Dispase, Corning Cell recovery solution or Accutase for recovering enteroids from Matrigel? I would appreciate any suggestion you may have.
Hi! I am not sure which protocol you are using apart from those compounds, but you can add 5mM EDTA (dissolved in PBS) and rock the samples at 4C for one hour. This should dissolve the drops of Matrigel. I have been using this method to remove the matrigel from my human gastrointestinal enteroids and it works great.
I would add that when you dislodge the drops of matrigel from your plates, use a 1mL pipette to start breaking the matrigel (be sure this doesn't break your cow enteroids) then I would add no more than 3-4 drops of matrigel per 15mL tube of EDTA solution. If you have a lot of matrigel it is not going to dissolve well and you are going to have it there still.
I think there is more info in our paper: Article Controlling Epithelial Polarity: A Human Enteroid Model for ...
good luck!
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