Immortalization of Epithelial Cells and Methods of Use
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Abstract
The present invention is directed towards methods of culturing non-keratinocyte epithelial cells, with the methods comprising culturing non-keratinocyte epithelial cells in the presence of feeder cells and a calcium-containing medium while inhibiting the activity of Rho kinase (ROCK) in the feeder cell, the non-keratinocyte epithelial cells or both during culturing.
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Citations
79 Claims
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1-58. -58. (canceled)
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59. A method of continuously culturing non-keratinocyte epithelial cells, the method comprising
a) culturing the non-keratinocyte epithelial cells in the presence of feeder cells and a calcium-containing medium, and b) inhibiting the activity of Rho kinase (ROCK) in the feeder cell, the non-keratinocyte epithelial cells or both during culturing.
- 69. A population of conditionally immortalized non-keratinocyte epithelial cells.
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73. A method of identifying a candidate treatment for a subject in need of treatment of a condition that is marked by the presence of abnormal non-keratinocyte epithelial (NKE) cells, the method comprising
a) obtaining a sample of the abnormal NKE cells from the subject, b) culturing the abnormal NKE cells in the presence of feeder cells, a calcium-containing medium and at least one Rho kinase (ROCK) inhibitor, to produce a population of abnormal NKE cells in vitro, c) determining a response profile of at least a portion of the abnormal NKE cells in vitro, and d) identifying a candidate treatment for the subject based on the determined response profile.
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77. A method of identifying an abnormal non-keratinocyte epithelial (NKE) cell in a subject, the method comprising
a) culturing at least one candidate abnormal NKE cell isolated from the subject in the presence of feeder cells, a calcium-containing medium and at least one Rho kinase (ROCK) inhibitor, to produce a population of candidate abnormal NKE cells in vitro, b) determining a tissue origin profile of at least a portion of the population of candidate abnormal NKE cells in vitro, and c) comparing at least one feature of the candidate abnormal NKE cells to the same feature of normal NKE cells that are obtained from the same tissue as that of the determined tissue origin profile of the candidate abnormal NKE cells to determine if there is a difference between the candidate abnormal NKE cells and the normal NKE cells, d) wherein a difference indicates that the candidate abnormal NKE cells are abnormal compared to normal NKE cells.
Specification