Proliferation/Differentiation

Resources
HGEP

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.

General Thawing, Cultivation and Freezing Protocol


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.

Aging in CnT-AG2

3D_Epidermis_sml

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.

2D Keratinocyte Differentiation Protocol

3D Keratinocyte Starter Kit – with HPEK Keratinocytes

3D Keratinocyte Starter Kit – without HPEK 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.

Video: Establishment of 3D epidermal cultures (12 MB, .wmv). Please see written protocols for description of process improvements.

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

Protocol for Establishment of Full Thickness Skin Models

msc_p4_defined_lipids

Differentiation of MSCs

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.

Adipogenic Differentiation of MSCs

Osteogenic Differentiation of MSCs

Chondrogenic Differentiation of MSCs

Airway3D_Day20

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.

2D Differentiation of Large Airway Epithelial Cells

3D Differentiation of Large Airway Epithelial Cells

Melano1

Melanocyte Differentiation and Total Melanin Assay

Differentiation-optimized media are recommended when melanocytes are induced to express more melanin.

For the differentiation of melanocytes and a method for evaluating total melanin, please see the following protocol.

Melanocyte Differentiation