Corneal Epithelium Culture Media

CELLnTEC has the following specialised corneal culture media available:

CnT-20

As a result of its super-optimized nutrient profile and PCT formulation, this fully defined medium offers both excellent isolation and extended growth of human corneal epithelium.

This defined medium can even equal or outperform traditional non-defined media, for example those containing Bovine Pituitary Extract or feeder layers.

In fact, a study done at Baylor College in Houston Texas (presented at ARVO 2007) showed that this medium provides better isolation, growth, and stem cell marker expression than alternative media from Invitrogen, Cambrex, and Cascade Biologics.

The PCT formulation of CnT-20 is known to delay the onset of differentiation. Thus for experiments where cells are induced to differentiate, the culture should be switched to CnT-30 24-hours prior to the induction of differentiation.

Seeding density is known to affect the performance of corneal cell isolation and growth in vitro. For this medium, good isolation and growth have been found to occur at densities above 4x10^3 cells per square cm, with optimal results at 8x10^3 cells per square cm.

CnT-50

For the absolute best isolation and growth, high colony forming efficiency and extended longevity, CnT-50 combines all the progenitor cell retention benefits of a PCT medium with the isolation and growth boost of BPE.

In the following experiment, CnT-20 and CnT-50 were compared with KSFM (Invitrogen) and SHEM (containing FCS) for their isolation ability.

As evident from these pictures, both CnT-20 and CnT-50 establish significantly more cells following isolation. For more comparisons over additional passages, please click here.

An additional benefit of CnT-50 is its low-BPE formulation. Traditional epithelial culture media typically use high BPE concentrations (e.g. Invitrogen and Cambrex media falling in the range of 25-50 ug/mL). In contrast, the optimum concentration of BPE in the PCT CnT-50 has been found to be only 15 ug/mL. Thus this medium provides all the benefits of BPE whilst minimizing any potential variability and the influence of unwanted factors.

For optimal results, we recommend seeding 4x10^3 cells per square cm at isolation, and 2x10^3 cells per square cm for passaging when using this medium.

CnT-30

Formulated with an optimal nutrient profile, CnT-30 provides good growth of corneal epithelium, and should be used in any experiments where cells are induced to differentiate.

This medium does not contain PCT factors, and is low in calcium (Ca2+).

Cells should be switched from CnT-20 or CnT-50 into CnT-30 24 hours prior to the induction of differentiation by the addition calcium (or your normal protocol). See our Resources Page for a suggested protocol.