Hello guys,
I am a beginner in biology, I wanna ask you about a question in cell culture.
I have cultured 1 - 2 mm fiber with structure as hydrogel shell, and HUVEC and collagen core in HUVEC media.
After 5 days, HUVEC are growth and change the morphology, form the monolayer as well as occur sprout angiogenesis process
However, after that (day 6), I cannot observe any new tube of sprouting (disappear), and all HUVEC in the collagen matrix was moved from 3D fiber to dishes.
This is the first time I have seen it, I tried to figure out but I can't.
If you have any answer or ideas, please feel free to reply.
How do I prevent this phenomenon?
Thanks a lot!
Take care!
Dear Alex
You can culture the HUVEC cells in a 2layer collagen hydrogel in which the inner layer made from the collagen with the less concentration. Using this culture system the cells will maintain their ability to migrate while will be entrapped in the denser 3D collagen hydrogel surrounding the inner one.
Dear Alex Nguyen,
From our experience on Matrigel, the tube formation from HUVEC culture is transient and seldomly to maintain for a long time due to its native property I guess. Thus, what I could say that you need to identify the specific time frame for your study so that you could trace and record the tube formation. In addition, you could try Alireza suggestion but you need to see your microscopy system ability to view and record the cells in 3D (a limitation specifically the observation distance between your lense and sample). Best of luck!
Hi, dear Fauzi,
I am also having the same problem. We also observed after a few days, the cells migrated from the 3D surface to TCPS. In order to achieve a 3d model, you need to maintain optimal cell population in the scaffold. I suggest using a hydrophobic surface to passivate the cells from moving from the plates to the scaffold.
All the best
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