I've been using Growth factor-reduced Matrigel for spheroid invasion for almost a year now and I didn't have any problems. However, recently I started encountering some problems.
In most cases, I see a few cells invading on the second day (as shown in the picture on the right). However, lately, in addition to "darker" invading cells, there are also those single invading cells that seem to detach from the spheroid more easily (as shown in the picture on the left). So far, I have tried varying the time that I keep the plate on ice before the addition of Matrigel (a few minutes up to 10 minutes) and whether I keep the plate on ice after the addition of Matrigel (none to 3 minutes). These are all the same cells seeded at the same density and Matrigel concentration is consistent).
Has anybody encountered a similar thing? Any suggestions are more than welcome.
Are you using a new batch of Matrigel? Even the reduced growth factor product vsries in growth factor content from batch to batch. You may need to ask for test samples and select the batch in which your spheroid cells behave as they did before.
Dear Ellen A G Chernoff thank you for your answer.
I checked and it was the same batch. But I did figure out that I've used different media in these two cases and the one on the left had growth factors added.
Also, the one on the left has 1%, whereas the one on the right has 10% FBS (I don't know if that could contribute to this).
Absolutel. The high growth factor and adhesive glycoprotein content at 10%FBS will affect cell migration. The cells could also be nutrient limited at the 1% level.
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