I have only worked with immortalized and tumoral cells. Now I have to separate epithelial cells from human tissues.
What type of tissue do you want to isolate your primary cells from? I work with primary human skin cells.
The most easiest way to get normal epithelial cells is ascites. Ascites is a common finding in patients with e.g. ovarian cancer. The problem with normal epithelial cells is that the cells have limited division capacity. You can split them at most 5 times, then the cells show all signs of senescence.
Thank you for your answers. The specific tissue is human endometrium. I have read some works in which is shown the specific growth medium, but I would need more technical details ( ex. cell strainer, contamination). I know the problem of limited division capacity. Is it better to immortalize the cells or avoid further modifications?
Generally, endothelial, epithelial or mesenchymal cells are perfectly growing (for 5 passages) in Iscove medium with 8% FCS (not "inactivated!!), and glutamin, Pen,Strep as usual. Contaminations are usually not a problem. Dissociate the tissue with collagenase. I don't know what you want to do with your cells but in any case I would avoid artificial immortalisations since the cells change incalculably. Maybe an alternative is a tumor cell line of your desired tissue. Endometrial carcinoma or sarcoma are not rare. But this depends on your experiments.
You can look up the materials and methods section of the following paper:
Lecanda J, Ganapathy V, D'Aquino-Ardalan C, Evans B, Cadacio C, Ayala A, Gold
LI. TGFbeta prevents proteasomal degradation of the cyclin-dependent kinase
inhibitor p27kip1 for cell cycle arrest. Cell Cycle. 2009 Mar 1;8(5):742-56. Epub
2009 Mar 16.
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