Establishment and characterization of a sheep endometrial epithelial cell line.

Endometrial epithelial cells play a significant role in the "dialogue" between the embryo and the mother, but in vitro studies to clarify this are hampered by the limited lifespan of primary cells. As such, it is necessary to develop an in vitro model to study endometrial function. Morphological analysis showed that the pEECs were homogeneous, formed characteristic cobblestone monolayers, and expressed the epithelial cell-specific marker, cytokeratin 18. The isolated and purified cells were transfected with a plasmid encoding human telomerase reverse transcriptase (TERT) gene, pCI-neo-TERT, to establish an immortal endometrial epithelial cell line (iEECs). The transfected cells were cultured with G418 and monoclonal cells were selected for expanded culture. Expression of TERT mRNA was detected by RT-qPCR and protein was quantitated by Western blot. TERT expression was stable and continued to be active with no signs of aging. Assays for cell proliferation and apoptosis indicated higher proliferation and cellular activity in iEECs than pEECs. After stimulated by interferon tau (IFN-τ), both iEECs and pEECs showed similar upregulation levels in all the underlying genes. Taken together, these findings demonstrate that iEECs retained the basic morphology and function of pEECs, providing a robust in vitro model for study of the function of ovine endometrial epithelial cells.

A pilot study on biological characteristics of human CD24(+) stem cells from the apical papilla.

BACKGROUND/PURPOSE: CD24 is a specific cell surface marker for undifferentiated dental stem cells from apical papilla (SCAPs) seen only during root development, before the tooth emerges through gum. But the comprehensive role of CD24 in the SCAPs is unclear. This study aims to clarify the exact roles of CD24 in SCAPs. MATERIALS AND METHODS: SCAPs were divided into CD24 (+)-SCAPs (high percentage CD24) and CD24 (-)-SCAPs (low percentage CD24) via flow cytometry. The proliferation, migration and osteogenic/adipogenic differentiation of the two groups were detected, RT-PCR was performed to detect the expression of osteogenic/adipogenic related genes and thegene expression were analyzed. RESULTS: The proliferative and migratory ability of CD24 (-)-SCAPs were significantly stronger than that of CD24 (+)-SCAPs. Although, the mineralization process and the osteogenic genes expression were not significantly difference in the two groups. Both CD24 (+)-SCAPs and CD24 (-)-SCAPs differentiated into adipocytes. The adipogenic differentiation in CD24 (+)-SCAPs was better than that in CD24 (-)-SCAPs, after 3 weeks of adipogenic induction. However, the expression of adipogenic related gene, PPAR γ2 mRNA in CD24 (+)-SCAPs was lower than that in CD24 (-)-SCAPs after 1 week of adipogenic induction. But the trend changed for the opposite after 3 weeks. CONCLUSION: The study proposes that CD24 has a regulatory effect on the adipogenic differentiation of SCAPs, and this may be attained by targeting the PPAR γ2 mRNA. Concurrently, it was found that CD24 plays an inhibitory role in the proliferation and migration of SCAPs, which may minimize the manifestation of diseases caused by an abnormal cell growth.

Effect of miR-204 in cell biological characteristics of breast cancer MDA-MB-231 cell / 重庆医学

Objective To study the effect of microRNA-204 (miR-204) on the biological characteristics of breast cancer cells.Methods Real-time PCR was used to detect the expression of miR-204 in human breast cancer cell MDA-MB-231 after transfection of miR-204 mimics and inhibitor for 48 h.Flow cytometry was used to analyse the effect of miR-204 on the proliferation and apoptosis of MDA-MB-231 cells.The effect of miR-204 on the migration of MDA-MB-231 cells was detected by Transwell migration assay.Results Real-time PCR analysis showed that miR-204 mimics and inhibitors had significant effect compared with normal control group(P<0.01).Flow cytometry analysis showed that compared with normal control group,the number of G1 phase cells of miR-204 mimics group was significantly decreased(P<0.01),while the number of G2/M cells of miR-204 mimics group was significantly increased(P<0.01).In contrast,the number of G1 phase cells of miR-204 inhibitor group was significantly increased(P<0.01),while the number of G2/M cells of miR-204 inhibitor group was significantly decreased(P<0.01).miR-204 mimics group significantly promoted apoptosis,while the inhibitor group significantly inhibited apoptosis(P<0.01).Transwell migration analysis showed that the number of cells of miR-204 mimics group were significantly reduced,while the number of cells was significantly increased in the inhibitor group(P<0.01).Conclusion We find miR-204,which can promote cell apoptosis and inhibit cell proliferation and migration,is a negative factor in the breast cancer cell line MDA-MB-231.