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When subculturing proliferating HUVEC (T25 flask; C‑12250), which tissue‑culture flask should I use?

A subconfluent T25 flask of proliferating HUVEC typically contains approximately 0.9–1.2 million cells, corresponding to about 36,000 - 48,000 cells/cm².
 
Recommended approach
PromoCell recommends counting the cells after trypsinization, and calculating the required number and size of new flasks based on the recommended seeding density.
 
Seeding density
Recommended seeding density for HUVEC: 5,000 - 10,000 cells/cm²
 
Typical split ratios
This seeding density usually corresponds to a split ratio of 1:4 to 1:6.
1:6 split ratio means that the total culture surface can be increased by a factor of 6, for example:
  • from 1 × T25 to 6 × T25, or
  • from 1 × T25 to 2 × T75
The exact choice of flask size depends on your experimental needs and available culture capacity.

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