Routine 2D Culture of CELLnTEC Primary Cells

Primary cells are sensitive, and can easily perform poorly without attention to the details or thawing, seeding, passaging and freezing. We recommend the following protocol for our full range of primary cells, in conjunction with specific info in the culture medium datasheet.

Aging of Keratinocytes with VitroAge Medium

Keratinocytes can be aged during several weeks of culture in VitroAge (CnT-AG2).

There are several important points to ensure dependable cell performance The following protoocol is recommended.


2D and 3D Differentiation: Epidermal Keratinocytes

Media designed for proliferation will always limit differentiation.

As a result, the following protocols are recommended for 2D and 3D differentiation of epidermal keratinocytes.

The following video is also available to illustrate the process for establishing 3D keratinocyte cultures.

Please be advised that this video was produced for our first generation 3D keratinocyte kits. Several elements of the process have since been improved.

Key changes include the culture media (now the CnT-Prime and 3D Barrier media), medium volume per insert and medium change frequency during airlift, and the faster establishment of the models (now day 12). Full details of the improvements can be found in the written protocols above.

FT Model

Full Thickness Skin Models: 3D Co-Culture

Full thickness 3D skin models can now be established routinely in your own lab, without the need for costly, complicated and variable collagen scaffolds.

The models are established using the new CnT-PR-FTAL medium, in which fibroblasts secrete all the ECM required for the dermal part of the model. For full details, please see the following protocol


Differentiation and CFE of Adipose-Derived MSCs

Adipose-derived MSCs growing in one of the CnT-Prime MSC expansion media retain the ability to differentiate for many passages.

To induce differentiation, the following protocols are recommended.

The percentage of active progenitor cells in an MSC culture can be evaluated using a colony forming efficiency (CFE) assay.

The following CFE protocol is recommended.


2D and 3D Differentiation: Large Airway Epithelial Cells

The use of differentiation-optimized media is recommended when inducing cells to differentiate.

For the routine 2D or 3D differentiation of airway epithelial cells, the following protocols are recommended.