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Showing 2 results for Co-Culture

Fatemeh Rezayat, Mehri Hajiaghaei, Dr Nariman Mosaffa, Dr Fahimeh Ramezani Tehrani, Dr Mehrnaz Mesdaghi,
Volume 18, Issue 2 (7-2016)
Abstract

Introduction: Polycystic ovary Syndrome (PCOS) is one of the most prevalent endocrinology disorders in women, in whom the state of systemic inflammatory cytokines, especially TNF-α is the main reason for immunological disturbances. Some PCOS manifestations such as infertility, hyperandrogenism, obesity and chronic inflammation are considered as risk factors for breast cancer. The risk of developing breast cancer in women with PCOS is being investigated in some epidemiological studies. In this research, the ability of peripheral blood mononuclear cells (PBMCs) of women with PCOS to develop antitumor response was studied and evaluated using an experimental co-culture approach between PBMCs and breast tumor cell lines. Materials and Methods: PBMCs were isolated from 50 heparinized venous blood samples (patient and healthy groups) by density gradient centrifugation byficoll. Breast cancer cell lines (MDA-MB-468 and MCF-7) were incubated as two target cells and were cultured adjacent to PBMCs in a transwell co-culture system. At different time intervals (48 and 72 hours) after co-culture, the proliferation rate of the effectors cells was evaluated by the BrdU cell proliferation assay. Determination of T CD3+CD8+ lymphocytes was determined by flow cytometry. Results: The proliferation of PBMCs after 48 hours of co-culture with MDA-468 (P=0.002) and MCF-7 (P=0.021) was significantly higher in the PCOS group compared to healthy controls. No pronounced differences were observed in T CD3+CD8+ cell numbers between the PCOS group and healthy controls (P>0.05) although T CD3+CD8+ percentage increased after 72 hours of co-culture in most samples. There was no statistically significant difference between MDA-468 and MCF-7 co-cultures in any of the tests. Conclusion: The stimulation threshold for mononuclear cells was reduced in women with PCOS. Differences between proliferation responses of PCOS and control groups may be caused by a chronic inflammatory condition in these patients.


Mehri Hajiaghayi, Fatemeh Rezayat, Nazanin Ghasemi, Amin Soleymanian, Mehrnaz Mesdaghi, Dr Nariman Mosaffa, Dr Fahimeh Ramezani Tehrani,
Volume 18, Issue 3 (9-2016)
Abstract

Introduction: Polycystic ovarian syndrome (PCOS) is a proinflammatory state that underpins the development of metabolic aberration and ovarian dysfunction in the disorder. Chronic inflammation and increased levels of androgens in these patients and their impact on the immune system, may be able to disrupt antitumor activity and thus increase the risk of developing malignancies including ovarian cancer. Materials and Methods: Peripheral blood mononuclear cells of 50 patients with PCOS and healthy controls were purified by Ficoll density gradient centrifugation. We then measured cell proliferation and concentrations of cytokines TNF-α at different time intervals (48 and 72 hours) after co-culture of ovarian (SKOV3, A2780) and breast (MCF-7, MDA-468) tumor cell lines with PBMC in indirect contact of transwell system. Results: Proliferative response of executive cells during stimulation with tumor cell lines after 48 hours was not statistically significant between patients and healthy controls. Between the 2 groups,  proliferation rates at the end of 72h were significantly higher than after 48h (P<0.01). The production of TNF-α in co-culture of A2780 cell lines significantly increased in the patient group in time compared to the controls (P<0.05). Conclusion: Our findings confirmed that, compared to healthy individuals low levels of chronic inflammation in patients with PCOS exhibit increased proliferative response of immune cells and TNF-α levels. An increased risk of cancers in patients with PCOS however requires investigation of other aspects of anti-tumor responses in vitro, with larger sample sizes.



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مجله ی غدد درون‌ریز و متابولیسم ایران، دو ماهنامه  پژوهشی مرکز تحقیقات غدد درون‌ریز و متابولیسم، Iranian Journal of Endocrinology and Metabolism
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