2D Differentiation of Epithelial Cells, Fibroblasts, and Melanocytes With Ease and Accuracy

2D Differentiation of Epithelial Cells, Fibroblasts, and Melanocytes With Ease and Accuracy

Conventional 2D platforms are attractive to cell biologists due to their simplicity and efficiency. 2D monolayer cell culture enables steady nutrients and growth factors supply which results in homogenous growth and proliferation.

The process of 2D differentiation is a highly orchestrated series of complex events that are significantly affected by the cell’s environment. A wide range of factors in the proliferation media actively counteract the cell differentiation process. Therefore, it is recommended to use media that were specifically developed for differentiation, such as CELLnTEC’s 2D differentiation media. These media come in a fully supplemented, one-bottle ready-to-use formulation designed to create an environment conducive to cell differentiation. To culture cells prior to differentiation, we highly recommend using our respective proliferation media to ensure optimal performance. 

Tissue TypeProliferation Medium2D Differentiation Medium
KeratinocytesEpithelial Proliferation CnT-PREpithelial Differentiation CnT-PR-D
FibroblastsFibroblast Proliferation CnT-PR-FFibroblast ECM CnT-PR-ECM
MelanocytesMelanocyte Proliferation CnT-40Melanocyte Differentiation CnT-PR-MD
Airway EpithelialEpithelial Proliferation CnT-PR-AMelanocyte Differentiation CnT-PR-AD


CELLnTEC’s Prime Epithelial 2D Differentiation (CnT-PR-D) medium delivers complete differentiation of primary epidermal keratinocytes and most other epithelial cells, while large airway epithelial cells can be differentiated using our CnT-Prime Airway Epithelial 2D/3D Differentiation Medium (CnT-PR-AD).  

  • Chemically defined, contains no components of animal or human origin
  • Optimal low-calcium formulation to trigger efficient differentiation by adding calcium at the desired time point

Protocols: [Keratinocyte 2D Differentiation]  [Airway Epithelial Differentiation]    

Application: McDew-White et al., 2021 induced cellular differentiation in primary human gingival epithelial cells using CnT-PR-D medium. Liang et al., 2022 differentiated human epidermal keratinocyte progenitors (HPEKs) and mouse epidermal keratinocyte progenitors (MPEK-BL6) using CnT-PR-D.

Keratinocytes differentiation with calcium

Following calcium switch, keratinocytes increase the expression of a wide range of differentiation markers. MPEK long-term mouse keratinocytes grown on CnT-PR-D, stained for cadherin adhesion molecules. Expression of the differentiation marker DSG3 (green) is evident at the cell borders following 5 days of differentiation.


Our Prime Fibroblast ECM (CnT-PR-ECM) medium induces ECM production in fibroblasts. CnT-PR-ECM is designed to have a less proliferative signal as cells undergoing rapid proliferation are less responsive to differentiation and ECM-related triggers.

  • Chemically defined, contains no components of animal or human origin
  • Encourages high secretion of extracellular matrix proteins from stromal cells

Protocols: [Fibroblast Differentiation]  

Application: Kolundzic et al., 2021 demonstrated a protocol to produce basement membrane substitutes using the CnT-PR-ECM medium. Coutier et al., 2022 established a protocol to generate dermal-epidermal constructs wherein the dermal equivalents were cultured with the CnT-PR-ECM medium.

Fibroblasts differention and ECM production

CnT-PR-ECM enables cells to produce in-vivo-like quantities of ECM because of its altered growth factor mix which shifts the focus away from strong proliferation, and the presence of vitamin C. Fibroblasts growing in CnT-PR-ECM deliver 5x higher ECM levels than proliferative cells growing in CnT-PR-F.


CELLnTEC’s Prime Melanocyte Differentiation (CnT-PR-MD) medium is designed for the differentiation of melanocytes and the production of higher levels of melanin. Melanocytes cultured in the CnT-PR-MD medium are less proliferative and display a much more in vivo-like phenotype.

  • Delivers increased dendricity of primary human melanocytes
  • Supports increased melanin production

Protocols: [Melanocyte Differentiation]  

Application: Crawford et al., 2020 used the CnT-PR-MD medium for the differentiation of murine epidermal melanocytes.

Melanocytes differentiation and melanin production

Differentiation and melanin expression is increased by 2.5x after melanocytes are switched to the CnT-PR-MD medium.

Originally posted by CELLnTEC on: https://us1.campaign-archive.com/?e=[UNIQID]&u=091fa9f299e4360f501d57555&id=a9ce9994d7

Caltag Medsystems is the distributor of CELLnTEC products in the UK and Ireland. If you have any questions about these products, please contact us.

2D Differentiation of Epithelial Cells, Fibroblasts, and Melanocytes With Ease and Accuracy
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