MatriGel plating - (Mar/21/2007 )

My PI told me to start using matrigel although nobody in the lab has used it previously so I have 2 questions: can you do a typical IHC on matrigel? I mean, do Antibodies have access to the cells? And can you visualize cells under a normal or fluorescence microscope or do you need a confocal? (I hope that no)

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Hello Pumuki!

I am not a specialist in this area but some consideration from my mind

Matrigel is a mixture of connective tissue proteins and cells seeding on this matrix can get signal for differentiation. To start cell differentiation is one of aims of matrigel as a layer for cells. So I don|t know, but may be some interferation with real IHC picture ( i mean morphology) because of this process.
I think that no problem with antibody access to the cell because of it is layer on what cells grown . Also I think you can visualize cells under a normal or fluorescence microscope.

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we have done some IHC on cells grown with matrigel as substrate and didnt have any problems.

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you may have no problems with ICC/IHC of Matrigel-cultured cells/tissues if you do not plate Matrigel too thick

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To all who use matrigel, check with BD to see if the lot you are using has been contaminated. Many recent kinds of Matrigel have come up positive for the LDEV virus. You may not be able to get clean matrigel for a year, at least.
Anyone know of any substitutes to matrigel?

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Thank you Jazen for this info!

According composition of Matrigel ( collagenIV type 31%, laminine 56%, entactin 8%) I think you can substitude with Collagen IV type or laminine or fibronectin. But it will be too expensive and depends on your tasks( differentiation or other exp) Sometimes collage I type from rat tails (cheap) is good essentially for adhesive assays but not for differentiation studies. I don't no other complex mixture of connective tissue proteins

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thanks Jazen for the info.

by the way where did you get this info about the matrigel.

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BD sent out a memo to all their customers of large volumes and extensive use.
I can post it on this site if you wish. Could have serious impact on in vivo studies, really doesn't effect anything in culture.

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Hello all,

I am lost. I don't understand why we should coat the plates with matrigel and what type of cells need that? I have grown some cells for animal studies. The day of injection, I trypsinized the cells and mixed cell suspension + Matrigel for injection. I handled Matrigel exactly the same way in this discussion - on ice, e.t.c. and that project was successful. But I don't understand this discussion. Just interested.
Thanks.

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thanks once again Jazen. I checked the BD website and didnt find any thing about matrigel so was wondering where you got the information. So I asked a sales rep and got a mail saying that ours was not contaminated. Anyway we use it only for cell culture.

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