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Can I add an additional layer of matrix proteins e.g. laminin or fibronectin at the end of the RAFT™ Process?

It is possible to add a layer of laminin or fibronectin at the end of an experiment after the RAFT™ Process has taken place. However, if cells have been embedded in the RAFT™ Culture, it is important to make sure that the culture is not left to dry...

What’s the difference in the immunofluorescence protocol for RAFT™ 3D Cultures and for 2 dimensional cell cultures?

First of all, immunofluorescence labeling works for RAFT™ 3D Cultures like for 2D cultures. The labeling protocol is basically the same. When visualizing structures within cellular aggregates such as tumoroids, a more intensive permeabilization step...

Which 24-well plates can be used with the RAFT™ 24-well Bundle Kit?

To ensure optimum 3D cell culture production, the use of 24-well plates (tissue culture treated) from Corning, Greiner, Millipore and Nunc is recommended. 24-well plates (tissue culture treated) from other manufacturers may be used, however these...

Can I add additional matrix proteins such as laminin and collagen type IV into a RAFT™ Culture with the addition of some basement membrane extract such as Matrigel™ or Geltrex™?

We have tested adding up to 5%v/v Matrigel™ into the RAFT™ Collagen Mixture. It should be noted that this requires that the absorbers be left in contact with the hydrogel for 30 minutes instead of 15 minutes (step 5 of the standard protocol). All...

Can I remove the RAFT™ 3D Cell Cultures from the plate in which they have been made?

This is easily done using a pair of forceps. First, pass the forceps around the edge of the culture to help dislodge it from the bottom of the well, and then pinch the edge of the culture lightly to avoid damaging the culture. When in contact with...

When a RAFT™ 3D Culture is subjected to an immunofluorescence labeling, should one expect a high background coming from antibody cross reactivity with the matrix?

Our Technical Note “Immunofluorescence Visualization of Fibroblast or Cancer Cells in RAFT™ 3D Cell Cultures” shows no immunofluorescence cross reactivity between the used antibodies and the rat collagen matrix, even with secondary antibodies...

When using the RAFT™ Culture System for embedding cells and having a layer of other cell type on top of it (example a cellular barrier model), is it possible to remove the top layer of cells later?

...assume that with a little careful handling, you should be able to re-suspend the overlaying cells off the collagen scaffold and the cells imbedded within the collagen RAFT™ Scaffold will remain intact in the well. In a second step you can use collagenase...
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