3d collagen culture - trouble

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anita.rocha
anita.rocha's picture
3d collagen culture - trouble

Hi,

I'm working with human foreskin fibroblasts (HFF1) in collagen 3D cultures (collagen solution type I, ultrapure, bovine from Sgma-aldrich) using 48 well plates.
I tried mixing different cell numbers in the collagen solution but after forming the gel (at 37oC for an hour) I add the celll medium and the next day the gel seems to have dissolved or contracted to a small, white, round chip.

Any idea to what I am doing wrong?

Happened to anyone? Any solution?

Thank you,

Anita

DasCopy
DasCopy's picture
Hi Anita,

Hi Anita,
 
I believe you just discovered a sort of historic phenomenon: It's that fibroblast really like to contract either collagen preparation. Check out papers about "collagen gel contraction" "hydrogel contraction" or "Collagen lattice contraction"... like this review here: PMID: 18638264
I've been to a seminar recently where some researchers made up a collagen gel including cells; they then compressed the gel so it would get stiffer... cells survied and the compressed gel was more stable. I'm more into collagen sponges, so I can't give you a protocol on gels, sorry for that.
 
good luck finding additional clues
 
Sven
 

swdbell
swdbell's picture
I have some question about

I have some question about angiogenesis method. I have a problem in experiment.
I do Spheroid assay for 3D spheroid by Hanging drop technique. Using matrix by collagen from rat tails so my matrix is not clear   I think it participate of collagen,   because it has a white fibrous whan see in well and debris in microscope.
From this problem impact for spheroid is not good,not circle,       cell of this expose form there in next time so don't see abnormal shape in first day.         I think this problem happens form collagen hasn't well of polymerization.
How can do it ?       I do collagen on ice 3-5 minitue.

Thanks you

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