Does anyone have an efficient way to reliably coat matrigel to glass coverslips?

Hello Mark -

We have found that hESCs do not attach particularly well to glass, even if coated. You may see clumps attaching in 3D structures without spreading. On the other hand, we have found that plastics from ibidi (ibidi.com), designed for imaging applications, do support normal hESC attachment and growth when coated with Matrigel. Try ibidi u-Dish 35mm, high, ibiTreat TC treated, for example. We used these plates in the attached publication.

Best,

Brigitte

Article Modulation of FOXD3 Activity in Human Embryonic Stem Cells D...

Brigitte L. Arduini

Hello Mark -

Best,

Brigitte

Article Modulation of FOXD3 Activity in Human Embryonic Stem Cells D...

Jonas Walter

Hi,

In my hands hiPSCs and derived NESCs are attaching nicely to matrigel coated glass coverslips, true the colonies sometimes do not look as good as on my standard nunc plates.

Another observation I made was that they didnt like a reduction in the concentration of Matrigel. So if you face some problems maybe try to increase the concentration.

Do you need more specific references?

Tobias A Beyer

Alternatively you can coat thermanox coverslips (http://www.thermoscientific.com/en/product/nunc-thermanox-coverslips.html) with matrigel. In my hands that seems to work quite well with H9 cells.

Mark James Davies

Thank you for your kind suggestions. Will give them a bash.

Tobias A Beyer

By the way, after staining the cells on the plastic, you can mount them on regular glass cover slips (not on microscope slides). Like that you can image through the glass and thereby avoid the autofluorescence problem of the plastic.

Tobi

Fjodor Merkuri

I know it has been a while since this question was first posted but I recently had to coat some glass coverslips with matrigel and thought I'd share my experience.

I essentially followed the CSH protocol below for washing and coating coverslips with poly-L-lysine. http://cshprotocols.cshlp.org/content/2008/5/pdb.prot4988.abstract

In short, I placed the coverslips in aqua regia (3:1 ratio of HNO3:HCl) for ~2 hours. Then proceeded to wash in double distilled water until the pH of the wash was ~5.5 and finally washed the coverslips in 70% EtOH and stored them (in 70% EtOH) in the cell culture hood. Once washed, I left the coverslips to dry in the hood and followed the poly-L-lysine treatment procedure outlined in the protocol.

The next day I coated the coverslips, placed inside plate wells, with Corning Matrigel (final concentration 0.5 mg/ml) for 1 hour at RT and dried for 30 minutes at 37C. My cells grew very well on the poly-L-lysine/matrigel coated coverslips but not on the coverslips that were just washed with aqua regia then coated in matrigel.

I did notice that, in one of my wells containing poly-L-lysine treated coverslips, the cells attached at first but then detached the net day. THis is strange since it is not consistent with the other wells that contained poly-L-lysine coated coverslips.

Hope this helps

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