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Characterization of a Three-Dimensional (3-D) Organotypic Skin Model using Keratinocytes and Mesenchymal Stem Cells Immortalized by hTERT

Green and black Keratinocyte-neonatal skin cells.

ASCB Meeting 2014

Philadelphia, Pennsylvania, United States

December 06, 2014


In this study, we compared primary keratinocytes (ATCC PCS-200-010) to hTERT immortalized keratinocytes (Ker-CT; ATCC CRL-4048), co-cultured with either primary fibroblasts (ATCC PCS-201-010), primary adipose-derived mesenchymal stem cells (MSCs; ATCC PCS-500-011), hTERT-immortalized fibroblasts (BJ5ta; ATCC CRL-4001), or hTERT-immortalized MSCs (hTERT-MSCs; ATCC SCRC-4000). We confirmed that both primary keratinocytes and Ker-CT are able to fully differentiate into skin equivalents in a 3-D culture model when co-cultured with primary fibroblasts, primary MSCs, BJ-5ta, or hTERT-MSCs. To confirm the functionality of the co-culture models, both the primary keratinocytes and the Ker-CT air-liquid interface (ALI) cocultures were subjected to a scratch assay. Re-epithelialization occurred in both cell lines, and interleukin 8 (IL-8) showed an increase in expression from day 0 to day 1 and 3, corresponding to migration of cells into the wound. The continuous nature of the Ker-CT cell line makes it an invaluable model for the research of keratinocyte biology, as it eliminates the issue of short life span and donor variation seen with primary cells.

Download the poster to explore the use of the Ker-CT cell line as a model for keratinocyte biology research.