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Analysis of Tissue Layer Stratification in Cells over Expressing Cdk4 and Long‐time Cultured Cells with Green Fluorescence Protein
Author(s) -
Terefe Eyuel Solomon,
Vaughan Melville Bailey
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.830.3
Subject(s) - involucrin , keratinocyte , keratin , microbiology and biotechnology , epithelium , staining , biology , laminin , basement membrane , pathology , cell culture , medicine , extracellular matrix , genetics , paleontology
I examined whether the stratification of keratinocytes, the primary cells of skin epithelium, that over expressed the Cdk4 and telomerase (hTER)‐which made them into immortal cells, had an influence on whether the keratinocytes tissue layer was organized as normal keratinocytes. These immortal keratinocytes were engineered to overexpress GFP so they could be used in wound healing models to monitor keratinocytes migration in real time as they healed a wound. The goal was to determine if GFP affected their ability to stratify and differentiate similar to normal keratinocytes. I arrived at my results by examining immunoflourcense stains for the presence of proteins within the epithelial layers/ dermal compartment in their proper location and orientation. The staining was for the proteins: Involucrin, Keratin 14, and basement membrane proteins collagen IV and laminin‐5. I found that the organization of the epithelial layers of the over expressing keratinocytes to be similar as those of non‐over expressing cells, which means that keratinocytes maintained epithelial cellular organization although they were going through abnormal proliferation. I also analyzed the epithelium layer organization of keratinocytes that were cultured for prolonged period. I also found that skin equivalents that were maintained for long period of time were similar to those that weren't maintained for such a long period. Grant Funding Source LS‐OKAMP