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Development of normal adult human colonic primary epithelial cell cultures and cell strains and their growth regulation - AN0312

Objective
1.Reproduce the conditions for the short-term normal human primary colonic epithelial cultures previously developed. Assess the normal primary cells for markers of epithelial cell differentiation such as keratins, CEA, E- cadherin, mucins and domeing. 2.Test a range of growth factors and dietary factors for their ability to act as survival factors and increase the long term survival and growth potential of primary colonic colonic epithelial cultures. 3.Test extracellular matrix components for their ability to act as survival factors and increase the long-term survival and growth potential of primary colonic epithelial cultures. 4.Examine the effects of growth/dietary factors on the expression of genes implicated in growth control and cell survival in primary cell cultures and determine whether changes in gene expression are related to the growth response of the cells. 5.Determine the effects of dietary short-chain fatty acids, calcium, and aspirin on proliferation, differentation and apoptosis on primary normal colonic epithelial cells to determine their possible role in maintaining normal tissue homeostasis. 6.Using a range of techniques described above (1-4) develop long term cultures and isolate colonic epithelial cell strains from normal adult colon and passage these cells in vitro to obtain frozen stocks. 7.Determine the role of short-chain fatty acids, calcium and aspirin in regulating proliferation, differentation and apoptosis in long-term epithelial normal cell strains.
Time-Scale and Cost
From: 1995

To: 1997

Cost: £78,315
Contractor / Funded Organisations
University - Bristol
Keywords