I am using the LONZA bullet kit media (CC-3162) kit for cultured HUVEC.
I have revived the cells 5 days back and cultured it in a two T25 flask. But till now I could observe only spherical shaped cells. I don't know why there is no change in the shape of the cells (elongated or cobblestone). However, the count of the cells is increased.
I have observed some of the cells are floating till now. I don't know why?
Could any suggest any solution to the above problems?
Hi Mariappan,
From your description it seems that you seeded HUVEC from one vial (frozen stock) in to 2 T25 flasks. That would suggest that the cell seeding density may be too low. My questions are: 1) How many frozen cells there were in your vial?, 2) Did you coat your T25 flasks with fibronectin, collagen, or gelatin before seeding HUVEC? (this is a critical factor for HUVEC to adhere/proliferate), 3) Did you change cell culture medium after the initial adhesion of HUVEC to the plastic (to remove DMSO that is present in the cell freezing medium)?
Overall, there may be different factors that keep your HUVEC "unhappy".... If HUVECs are seeded at the right density and are provided with the right cell culture medium (your LONZA kit seems to be OK), they should reach confluency in 4-5 days. HUVEC (if they are "true" endothelial cells) should not have "elongated" shape, unless are grown under specific (e.g. cell matrix-based) conditions.
Best
Gedas
- Badges
- Science topic
- Similar topics
- Computer Science and Engineering
- Computer Graphics and Animation
- 3D