I advise you to contact Prof. Ray Garry in Great Britain, [email protected].

Kind regards LILO METTLER from KIEL, Germany

[email protected]

You may know all of the info below. If so, I'm sorry. I thought I would give my perspective having been in this field for a while

First Question: Do you want epithelial lines or stromal?

Epithelium: There are a few normal cell lines out there, but most have not been extensively used, in part because their phenotypes seem unstable or because they don't grow well. HES cells, derived spontaneously from culture of primary human endometrial epithelial feels, have been extensively used (Desai NN et al. Fertil Steril 1994; 61:760–6) and may be available from Dr Kniss at ?Ohio State or Dr. Fazleabas at Michigan State Medical School or others. However, there is a rumor going around that most of the common endometrial epithelial cell lines (cancer and non-cancer) are actually either Ishikawa or HeLa.

Stroma: Primary stromal cells can be cultured for a number of passages, while maintaining a fairly differentiated state. Studies have observed differences between stromal cells taken from the uterine endometrium in women with versus without endometriosis and these differences seem to persist for a few passages (e.g. Aghajanova L, Hamilton A, Kwintkiewicz J, Vo KC, Giudice LC. Steroidogenic enzyme and key decidualization marker dysregula- tion in endometrial stromal cells from women with versus without endometriosis. Biol Reprod. 2009;80(1):105-114.). There is at least one transformed stromal cell line (temp sensitive T-antigen transfected?). Our experience (15 years ago?) with that line is that it did not grow well.

We do not have such cell lines. We assume that because endometrial epithelial and stromal cells respond to hormonal stimuli, properties of hybridomas thereof are likely to differ. One way to start with could be postmenopausla endometrium.

Thanks for the responses folks, Markku, my lab routinely prepares primary cells from human endoemtrium, but since these are short lived and not very homogeneous we have issues while doing some geentic manupualtions like overexpressions and knockdowns. Thus I would prefer some cell lines.

I need both steven, unfortunately , the only spontaneous epithelial cell line of endoemtrium is contaminated with HeLa and hence not to be used and there is no other option that i knew of.

thanks for sharing the info on the growth, I shall keep in mind

thanks Liselotte :)

Some points to ponder:

1. Cancer cells are referred to as immortal cells because of a transformed telomerase which allows them to escape senescence

2. Viruses transform normal cells(stem) capable of division to become immortal in vivo whereas normal cells have to be immortalized in vitro using e.g. SV40 T Antigen.

3. Normal endometrial cells are meant to be shed in a cyclic manner and that is why they lose the capacity to stick to any surface (in cultures) and immortal cells do not have the capacity to stick either.

4. Endometriosis is characterised by the overexpression of certain cell adhesion molecules and therefore metastasize and stick to all surfaces instead of being eliminated (also in culture)

5. Logically cell cultures made from normal endometrium would not stick whereas cells from patients of endometriosis will be able to stick.

6. Surface markers (CAMs) can be used to sort endometrial cells of patients from normal females by flow cytometry using antibodies.

7. Normal endometrial primary cells will not contain immortalized cells.

8. Normal endometrial cells populations should be sorted with flow cytometry immediately after preparation or after immortalization.

9. Endometriosis patients will yield transformed cells which will stick due to overexpression of CAMs but HeLa cells will not.

10. Refs for culture: Puerarin Suppresses Invasion and Vascularization of Endometriosis Tissue Stimulated by 17b-Estradiol. Dan Wang et al. PLoS ONE | 2011 | Volume 6 | Issue 9 | e25011

11. Refs for cell culture: L1 cell adhesion molecule (L1CAM) as a pathogenetic factor in endometriosis. D. Finas et al. Human Reproduction Vol.23, No.5 pp. 1053–1062, 2008

contd: points to ponder (I have not worked i this area)

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thanks so much manju for the papers. Did have a look at them.

I know about these 3D cultures and have been looking at them seriously :)

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