Related Experiment Video
Updated: Aug 7, 2026
Generating Primary Cultures for Keratinocyte Live Cell Imaging
Published on: May 2, 2025
Development of an optimised culture medium for keratocytes in monolayer
Nicolas Builles1, Nicolas Bechetoille, Virginie Justin
1Banque de Cornées des Hospices Civils de Lyon, 69437 Lyon, France.
Unlabelled:
Our objective was to formulate a medium for monolayer culture optimising both keratocyte growth and preservation of the keratocyte phenotype.
Methods:
An experimental matrix selected 14 media to test, using 7 components. Selection criteria were growth rates over 5 passages and expression of the CD34 marker.
Results:
Acetylcholine, insulin and vitamin C had no effect on growth and differentiation. The DMEM + Ham F12 1 : 1 based medium was selected for its initial effect on growth. At concentrations of 5 ng/ml, b-FGF improved the percentage of CD34+ cells without reducing growth rates. New-born calf serum (NCS) had a greater effect on growth than foetal calf serum (FCS). We showed three major interactions: between b-FGF and IGF-1, FCS and IGF-1 and NCS and b-FGF.
Conclusion:
We selected the following medium, which provides optimal growth and preservation of the CD34+ phenotype: DMEM/HAM-F12 + 10% NCS + 5 ng/ml b-FGF + antibiotics.
