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Quantitative characterisation of cell fate in human keratinocytes and squamous cell carcinoma

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  1. Tez No: 402715
  2. Yazar: GÖZDE AKDENİZ
  3. Danışmanlar: DR. PHIL JONES
  4. Tez Türü: Doktora
  5. Konular: Genetik, Genetics
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 2011
  8. Dil: İngilizce
  9. Üniversite: University of Cambridge
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Belirtilmemiş.
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 328

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Özet (Çeviri)

Squamous epithelia are constantly shed and need to be replaced throughout adult life. The balance between cell production and cell loss is therefore crucial. If too few cells are produced the tissue will fail, while overproduction of cells is a hallmark of cancer. It has been long held that normal human epidermis is maintained by two types of proliferative cell populations: long lived, self renewing stem cells and transit amplifying cells which differentiate after a few rounds of cell division. However, a recent study in mice proposed that the epidermis is maintained by a single population of progenitor cells with properties distinct from classical stem or transit amplifying cells, that generates progenitor and differentiated daughter cells with equal probability. To address the question of whether the murine model could be applied to human tissue I characterised the cell fate of human keratinocytes, squamous cell carcinoma (SCC) cell lines and freshly isolated tumour cells quantitatively in vitro. I also challenged cells with lapatinib, a dual tyrosine kinase inhibitor that targets epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor – 2. I have used quantitative cell fate analysis to show that the murine model is applicable to a subpopulation of human epidermal keratinocytes in vitro. In addition, I have shown that in clonal 2D culture SCC cell lines continue to proliferate but in a 3D organotypic raft culture system this is not the case; rather a proportion of cells stop proliferating and express differentiation markers. Primary human tumour cells in vitro differ from established cell lines in that in clonal 2D culture they contain cells that have ceased proliferating. Finally, I have demonstrated the effects of EGFR and HER2 inhibition on cell fate in human keratinocytes showing it slows down cell proliferation while inhibiting terminal differentiation. These results highlight the importance of quantitative studies in analysis of cell fate in vitro.

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