Matrigel

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steffybidim
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Matrigel

Hi!

I was interested in using Matrigel as a 3D matrix but there are a few questions I cannot answer so far. If some of you are using Matrigel you might be able to help me find some answers?!

First of all, it seems unlikely that Matrigel will ever be approved by FDA as a componant in an implant as it is derived from mouse sarcoma. Am I right ?

Also, I am wondering how long the Matrigel would stay "active" once gelled, especially the laminin. I doubt that such a gel could be stored a few months before use ?

Thanks to you who might have some answers!

Ivan Delgado
Ivan Delgado's picture
 

 
Hi steffybidim,
To answer your first question: there is no reason why the FDA would not approve Matrigel for an implant, even if comes from mouse sarcoma, if the submission provides indisputable proof that it is safe for human use. Yet, providing that kind of proof is very expensive, so my guess is that it is unlikely someone would go through the hassle of getting it approved. The classical way of looking at this is that there are alternative compounds that, while not as good as Matrigel, do a pretty good job so the extra expense of getting Matrigel approved outweighs the return on investment.
As for your second question it is kind of tricky to give an exact expiration date since it is not easy to quantify the viability of Matrigel's components. My educated guess is that once you've gelled Matrigel, it is only good for a few weeks tops. Remember that most Matrigel is only good for up to 3 months stored at -20oC, so once activated it will not last longer than that. 
 

steffybidim
steffybidim's picture
Thanks a lot Ivan!

Thanks a lot Ivan!

VBI
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Ivan,

Ivan,
I would imagine the reason for Matrigel not being worthy of approval for in human use is that it has to be immunogenic, and that is a big red flag in the eyes of the regulatory people.  The materials used for implants and cellular therapies need to be nonimmunogenic.  This is usually established very early in preliminary investigations.  With the push on regenerative medicine and stem cells gaining momentum, this will be a hot area.

Ivan Delgado
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Hi VBI,
I would not say that Matrigel is not "worthy", just hard to approve. And you are right, being immunogenic is definitely a problem hard to get around and the main reason why Matrigel likely will not get past the FDA.
 

minylim
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Hi Ivan,

Hi Ivan,

I have somewhat difficult problem with bioreactivity of matrigel. I found GFR matrigel often interfere with activity of growth factor of my interest. I've tested different lots but the expired, years-old lot gave best result. =( I'm seeking for the way to reduce the bioreactivity. I'm thinking of keeping solidified matrigel for a while before use it or repeat a few freeze-thaws. Do you think it'll also affect other ECM components such as laminin and collagen? Can I run Western blot with matrigel?? Thanks!!

samm
samm's picture
Have you ever tried a mix of

Have you ever tried a mix of 'inert' gelling agents like PhytaGel with different collagens? That will have minimal growth factors compared to MatriGel/Collagen mixes and might save you some time.

minylim
minylim's picture
Thank you for the suggestion

Thank you for the suggestion Samm. I'm about to graduate and my PI wants me to focus on 'producing' paper or data for grant renewal. I don't think my circumstance now allow me to try that but I'll definitely consider it  if I do more 3d culture in future.
Other question about collagen (I used type I and others in different labs had same trouble with same product) is it's quite soft and occasionally doesn't solidify. Did you ever have the problem? 

samm
samm's picture
Actually yes - and its not

Actually yes - and its not just you and me either! Thats why I use "collagen alone" only to coat (esp for primary cultures), and not as a relatively thick 3d scaffold. If you are interested in using collagen to make thicker coats, check out the protocol with bagged ammonia (!) that was posted in this site a few months back.
Also, you can run a Western with Matrigel (try a 8-16% gradient to get ~10-200 kD range) and probe to see the amounts of the growth factor in question, present in each batch you use. I'll try to dig up the papers where they had done just that - I think it was for ex vivo tumor cell culture.

helper
helper's picture
What type of Collagen I are

What type of Collagen I are you using (Human or Rat?)  - at what concentration are you trying to form a firm gel with it? 

minylim : 9/17/2009 10:29 AM
Than

k you for the suggestion Samm. I'm about to graduate and my PI wants me to focus on 'producing' paper or data for grant renewal. I don't think my circumstance now allow me to try that but I'll definitely consider it  if I do more 3d culture in future.
Other question about collagen (I used type I and others in different labs had same trouble with same product) is it's quite soft and occasionally doesn't solidify. Did you ever have the problem? 
 

minylim
minylim's picture
It was Rat Tail Collagen I

It was Rat Tail Collagen I from BD. I used different concentration (up to 100%) and tried different gellation methods (heat and pH, following the product manual), none of my condition worked. GFR matrigel got me best result even I've been suffered from lot to lot variation. =)

helper
helper's picture
It was Rat Tail Collagen I

It was Rat Tail Collagen I from BD. I used different concentration (up to 100%) and tried different gellation methods (heat and pH, following the product manual), none of my condition worked. GFR matrigel got me best result even I've been suffered from lot to lot variation. =)
 

Thats strange.  Rat Tail Collagen usually forms a nice gel even at a 1:10 dilution.  If your Collagen is old it would not gel properly, check that.  Are you using it as a gel or are you adding cells directly to the Collagen and trying to form a gel?  BD also offers a Laminin product that can be used to form gels.   

StudentLabrat
StudentLabrat's picture
I was searching the internet

I was searching the internet for information on the dangers of matrigel in humans, because as I was preparing pure matrigel (BD Biosciences) for control injections, one of my co-workers walked by very fast and shoved my arm and I actually don't know exactly how but I accidentally got stuck with the needle on my opposite arm, and since I was in the middle of getting the air out my finger was on the plunger about 50 ul of matrigel was injected into my arm!  I am terrified that I'll get cancer now since it is made from mouse sarcoma.  Has this been testing in humans?  Does it actually cause cancer?

Ivan Delgado
Ivan Delgado's picture
Unfortunately there is no

Unfortunately there is no conclusive answer to your question. As you likely know Matrigel is a trademark name for a proprietary mixture of components that is not fully understood and even varies from lot to lot.

As far as I know the only state that lists Matrigel as a cancer causing agent is California. Why I am not sure. Over 95% of what makes up Matrigel is Laminin, Collagen IV and Entactin, none of which are listed as cancer causing agents in California. Likewise none of the growth factors in Matrigel (bFGF, EGF, IGF-1, PDGF, NGF, and TGF-beta) are listed as cancer causing agents in California. 

For a complete list of cancer causing agents listed by proposition 65 in California go to this link.

Ultimately I cannot tell you that Matrigel is 100% safe. Obviously anything you get injected with in the lab is not good, but of all the possible accidents you could have experienced, this one does not seem to be one of the worst. My only recommendation is to call the company that sells the Matrigel you used and ask them if they have any opinions about your situation.

StudentLabrat
StudentLabrat's picture
Thanks Ivan, the first thing

Thanks Ivan, the first thing I did was look up the MSDS, which has the ingredient chloroform listed as a carcinogen, and also cell growth factors.  I actually did email the company (good advice) and hope they will reply to me Monday.  The good thing is that in all of my experiments, none of the immune compromised mice have developed tumors with the control matrigel vs. tumor + matrigel. 

The second thing I did was google the heck out of it for over two hours before I posted. I read every single piece of info, yes I know it's an extracellular matrix and all that stuff. Still there was no peer reviewed journal published research showing results in humans, just the phrase "NOT FOR HUMAN USE" over and over.

And being the only other person working on a Sunday afternoon, there was no one in the entire lab to report this to except for my dumb co-worker who doesn't know anything at all. And my boss is on vacation until the end of next week. And there is absolutely nothing the ER or my doctor can do, as it immediately congeals at body temperature and absorbs into the body, so I guess they could cut my arm off but I think that's a bit drastic.

Anyway, I emailed the company directly and am trying not to worry too much. If I grow a mouse leg or head out of my arm I'll let you know. :P

Ivan Delgado
Ivan Delgado's picture
It sounds like you've done

It sounds like you've done everything you can possibly do. I do not want to belittle this accident in any way, you always want to be very careful when working with these reagents, but my best guess is that you will be perfectly fine.

Hopefully the company will contact you back right away. If not, give them a call. In the mean time the best you can do is relax and not worry.

All the best!

Rns
Rns's picture
Hi Ivan,

Hi Ivan,
you wrote that over 95% of Matrigel is Laminin, Collagen IV and Entactin. I am doing 2DE for matrigel and wondered if you remember where you got that info from or if you can recommend me a paper detailing that info. 

Thanks,
Rens

Ivan Delgado
Ivan Delgado's picture
Hi Rens,

Hi Rens,

As far as I know you can get that information from a number of places. Here are a couple of links where that information is presented:

BD Matrigel matrix FAQ
BD Extracellular Matrix Proteins

Hope this helps,

Ivan