Profile of MicroRNA Expression in Endometrial Cell during In Vitro Culture According to Progesterone Concentration

Artificial uterus using endometrium implant can be a novel treatment strategy for infertile women with refractory endometrial dysfunction. At early pregnancy, the function of uterine endometrial cells for the communication between the conceptus of pre-implantation period and maternal reproductive system is essential. MicroRNA (miR) expression profile of endometrial cells according to progesterone, a crucial pregnancy-maintaining hormone, provides important data for in vitro endometrial cell culture strategy that is useful for engineering artificial uteri using endometrial implants. The present study aimed to evaluate the miR expression profile of in vitro cultured endometrial cells under hormonal milieu mimicking early pregnancy period in terms of progesterone concentration. We cultured murine uterine endometrial cells, human uterine endometrial carcinoma cells, and immortalized human uterine endometrial cells using different progesterone concentrations, and analyzed the expression of miRs critical for early pregnancy. The expression of miR-20a, -21, -196a, -199a, and -200a was differently regulated according to progesterone concentration in different endometrial cell lines. The analysis of candidate target genes showed that the expression of phosphatase and tensin homolog, mucin 1 (MUC1), progesterone receptor, transforming growth factor β receptor II, matrix metallopeptidase-9 was up-regulated by progesterone treatment in mouse and human endometrial cell lines. These results indicate that physiological concentration range (10⁻⁷ and 10⁻⁹ M) of progesterone affect the survival and target gene expression via modulating miR expression. Taken together, progesterone can be a crucial factor in regulating miR expression on in vitro cultured endometrial cells.

Effect of Taoren Quyu Decoction on human endometrial cells and its anti-endometriosis activity in rats

OBJECTIVE@#To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats.@*METHODS@#A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMAb), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected.@*RESULTS@#The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P  0.05).@*CONCLUSIONS@#TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.

Effect of Taoren Quyu Decoction on human endometrial cells and its anti-endometriosis activity in rats

Objective To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. Methods A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMAb), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected. Results The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P 0.05). Conclusions TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.

Interaction between endometrium and embryo under the condition of in vitro co-culture during the implantation window stage / 实用医学杂志

Objective To investigate the interaction between endometrium and embryo under the co-culture condition during the planting window stage. Methods Twenty patients who cancelled transplant for OHSS in the Reproductive Center of the second affiliated hospital of Zhengzhou university were enrolled.The discarded embryos were used to build the embryos co-culture system (Group A), with a single membrane (group B) and a single embryo(group C)as the control group. Levels of LIF and IGF expression and embryo growth were checked on 3, 4,5,6,7 day post-oocyte retrieval. Results Expressions of LIF mRNA and IGF mRNA in Group A were significantly higher than those in Group B. Embryo growth in the Group A was better than that in Group C. Conclusion Co-culture of endometrium and embryo supports the development of embryo and improves the receptivity of endometrium.

Clinical analysis of histopathology diagnosis of micro endometrial tissues obtained by endometrial cell collector / 中国医师进修杂志

Objective To evaluate the feasibility and application of histopathology diagnosis of endometrial tissues obtained by endometrial cell collector (Jingyou, SAP-1). Methods One hundred and ninety-three patients whose endometrial lesions should be excluded were selected. First, endometrial tissue were obtained from the patients by Jingyou, then they underwent comprehensive curettage under hysteroscopy. The histopathology diagnosis was performed respectively. The specimen satisfaction rate and diagnostic accuracy was analyzed and compared. Results The specimen satisfaction rate of curettage under hysteroscopy was 95.85%(185/193). The specimens of 8 cases were not satisfied because the tissues were not enough. The specimen satisfaction rate of Jingyou was 82.38% (159/193). The specimens of 34 cases were not satisfied, among whom in 10 cases scratches did not throughout the whole palace antrum, and in 24 cases tissues quality were poor. The endometrial thickness in unsatisfactory specimen by Jinyou was significantly thinner than that in satisfactory specimen:(0.64 ± 0.18) cm vs. (0.97 ± 0.43) cm, and there was statistical difference (P<0.05). The diagnostic accordance rate between Jingyou and curettage under hysteroscopy was 79.87%(127/159). The sensitivity of Jingyou from high to low was 94.19% (81/86) in normal menstrual endometrial, 7/10 in endometrial carcinoma/ atypical hyperplasia, 67.86%(38/56) in endometrial hyperplasia and 1/7 in endometrial polyps. Missed diagnosis of jingyou inluded 2 cases of endometrial carcinoma and 4 cases of endometrial atypical hyperplasia. The misdiagnosed rate of high grade endometrial lesions was 6/16, and the patients were misdiagnosed because the tissues were not enough. Four cases of endometrial atypical hyperplasia had underwent conservative treatment of repeated curettage. Conclusions Application of Jingyou can obtain micro endometrial tissues, and the accordance rate of histopathology diagnosis is high with curettage under hysteroscopy. When the collector makes a comprehensive collection to the uterine cavity specimen, it can accurately screen endometrial carcinoma and atypical hyperplasia. The patients who have endometrial atypical hyperplasia and receive conservative treatment and curettage repeatedly curettage, thin endometrium and ultrasonic highly suspected endometrial polyps, are not recommend to use Jingyou to obtain specimen. When the specimen is not satisfied using the collector, additional hysteroscopy should be performed to avoid misdiagnosis of high grade endometrial lesions.

Effect of active compound components of SQ on MMP-1,TIMP-1,VEGF,t-PA and PAI-1 of in? ammatory endometrial cell / 中华中医药杂志

Objective :To study the effect of MMP-1,TIMP-1,VEGF and t-PA,PAI-1 in the inflammatory process of endometrium cell and the active components of SQ.Methods: ICC,ISH and WB was used to test the level of protein of MMP-1,TIMP-1,VEGF and t-PA,PAI-1 and mRNA of MMP-1,TIMP-1 and VEGF of endometrium cell in regular group,pattern group,GongXueNing group and SQ group.Result: In contrast with model group,the level of MMP-1protein and mRNA expression obviously decreased,but the level of TIMP-1 and VEGF increased in SQ group,there were signifi cant differences(P

In vitro culture of human Endometrial cells / 医学研究生学报

Objective:To establish a method of endometrial glandular cells culture,storage and subculture in vitro,so as to study its antigen components.Methods: Endometrial glandular cells and stromal cells were isolated by collagenase digestion and copper sieve filtration methods and monolayer cultured. Results: Glandular cells and stromal cells in the culture had been observed by light microscopy and stained positively by keratin and vimentin McAb respectively using immunohistochemical method. The glandular cells life span can last 4-6 weeks and 2-3 population doublings. Conclusion: Our establishment of endometrial glandular cells show great promise for providing more non-hysterectomy origin EMAg.

Establishment of Three Dimensional in-vitro Culture System with Human Endometrial Cells: Induction of Differentiation by Sex Steroid Hormone & Characterization / 대한산부인과학회잡지

OBJECTIVE: The aim of this study was to establish three-dimensionally cultured endometrial cell model containing endometrial stromal cell (ESC), endometrial epithelial cell (EEC) and extracellular matrix (ECM) and to compare the morphological and biomolecular expression patterns of this model with mid-luteal endometrium in vivo. MATERIALS AND METHODS: The EEC and ESC was obtained from hysterectomy specimen and cultured separately. The EEC was overlayered in Matrigel layer on ESC embedded in collagen. The model had been cultured for 48 h in DMEM medium containing estrogen and progesterone. The ultrastructure was evaluated by electron microscopy. The expression of integrins, cyclooxygenases and matrix metalloproteinases were examined by immunohistochemistry and zymography. RESULTS: EEC in three-dimensional culture model grew with polarity and tight junction and desmosome between cells were found. The formation of pinopodes was also detected. In three-dimensionally cultured endometrial cell model, the expression of integrin alpha1, alpha4, beta3, MMP-1, -2, -3 and 9 was detected which was not expressed in monolayer culture of EEC, ESC or ESC embedded in collagen. CONCLUSION: The three-dimensionally cultured endometrial cell model possessed the morphological and biomolecular characteristics of in vivo endometrium of implantation period. These characteristics could be achieved by paracrine interactions between ESC and EEC. This model may contribute to the studies of differentiation of endometrium, process of implantation and pathophysiology of implantation-related diseases.

Inhibitory effect of Nitric oxide on the Proliferation of Endometrial cells / 대한산부인과학회잡지

OBJECTIVE: Development of endometriosis is associated with endometrial stromal cell proliferation. The effects of nitric oxide (NO) on the proliferation of endometrial cells and the effects of peritoneal fluid from women with and without endometriosis on the production of NO and NO-induced changes were investigated. METHODS: Endometrial stromal cells were prepared from women with endometriosis (n=10) and healthy control (n=10). Peritoneal fluid from 10 patients were collected at the beginning of laparoscopic procedures. The proliferation of cells was assessed by [3H]thymidine. Nitrite, a stable NO product, was measured with Griess reagents. Expression level of endothelial nitric oxide synthase (eNOS) was analyzed by quantitative RT-PCR. RESULTS: Sodium nitroprusside, an NO donor, reduced proliferation of the endometrial and endometriosis cells from primary culture in a dose-dependent manner. The increased production of NO and inhibitory effect of NO on the endometrial stromal cell proliferation were reduced by peritoneal fluid from patients with endometriosis but not by interleukin-8, 17-beta estradiol or TGF-beta1. Transcripts of the endothelial NOS (eNOS) gene were detected in unstimulated endometrial cells and the expression patterns were not changed by exposure of cells to SNP or peritoneal fluid for 24 hours. CONCLUSION: It is postulated that these findings show NO inhibits proliferation of endometrial cells and the inhibitory effect of NO on endometrial cells was abrogated by certain factor (s) of peritoneal fluid from patients with endometriosis. In conclusion, NO appears to be involved in the pathogenesis of endometriosis.