Endometrial stromal cells and decidualized stromal cells: Origins,transformation and functions

Decidualization of endometrium, which is characterized by endometrial stromal cell (ESC) decidualization, vascular reconstruction, immune cell recruitment, and plentiful molecule production, is a crucial step for uterus to become receptive for embryo. When implantation takes place, ESCs surround and directly interact with embryo. Decidualized stromal cells (DSCs) are of great importance in endometrial decidualization, having a broad function in regulating immune activity and vascular remodeling of uterus. DSCs are shown to have a higher metabolic level and looser cytoskeleton than ESCs. What's the origin of ESCs and how ESCs successfully transform into DSCs had puzzled scientists in the last decades. Breakthrough had been achieved recently, and many studies had elucidated some of the characters and functions of DSCs. However, several questions still remain unclear. This paper reviews current understanding of where ESCs come from and how ESCs differentiate into DSCs, summarizes some characters and functions of DSCs, analyzes current studies and their limitations and points out research areas that need further investigation.     

Two Methods for Establishing Primary Human Endometrial Stromal Cells from Hysterectomy Specimens

Many efforts have been devoted to establish in vitro cell culture systems. These systems are designed to model a vast number of in vivo processes. Cell culture systems arising from human endometrial samples are no exception. Applications range from normal cyclic physiological processes to endometrial pathologies such as gynecological cancers, infectious diseases, and reproductive deficiencies. Here, we provide two methods for establishing primary endometrial stromal cells from surgically resected endometrial hysterectomy specimens. The first method is referred to as “the scraping method” and incorporates mechanical scraping using surgical or razor blades whereas the second method is termed “the trypsin method.” This latter method uses the enzymatic activity of trypsin to promote the separation of cells and primary cell outgrowth. We illustrate step-by-step methodology through digital images and microscopy. We also provide examples for validating endometrial stromal cell lines via quantitative real time polymerase chain reactions (qPCR) and immunofluorescence (IF).     

Expression and hormonal regulation of calcyclin-binding protein (CacyBP) in the mouse uterus during early pregnancy

Calcyclin-binding protein (Siah-1-Interacting Protein, CacyBP/SIP), is a calcium signaling protein involved in the degradation of beta-catenin, however, little is known about its role in reproductive biology. The present study was to character its temporospatial expression pattern and regulation in mouse uterus and to investigate whether it plays a role in the regulation of normal endometrial events. While prominently expressed in both luminal and glandular epithelia, CacyBP underwent dynamic changes during early pregnancy. CacyBP expression was observed weakly from days 1-4. An intense accumulation in luminal and glandular epithelia as well as decidua surrounding the embryo at later stages (days 5-7) was observed. Most notably, CacyBP accumulation in trophoblast was pronounced at day 7. Using ovariectomized and pseudopregnant mice, we found that progesterone (P(4)) and 17beta-estradiol (E(2)) led to increased expression of CacyBP gene and this could be abolished by Ru486 and tamoxifen, respectively. Antisense oligonucleotides (ODNs) against CacyBP significantly inhibited cultured endometrial stromal cells' (ESCs) apoptosis induced by UV irradiation. Injection of antisense ODNs into mouse uterine horn severely impaired the number of implanted blastocysts. Taken together, our results suggested that CacyBP expression was positively regulated by P(4) and E(2). CacyBP may be involved in the regulation of endometrial cell apoptosis during early pregnancy and play an important role in mouse endometrial events such as pregrancy establishment.     

Effect of progesterone on embryo survival

Increased genetic selection over the past 40 years has resulted in a dairy cow with an improved biological efficiency for producing milk but with an associated reduced fertility. Embryo loss is the greatest factor contributing to the failure of a cow to conceive. The extent and timing of embryo loss indicates that 70% to 80% of this loss occurs in the first 2 weeks after artificial insemination (AI). This is the period when a number of critical phases in embryo development occur and where protein accretion, substrate utilization and embryo metabolism increase dramatically. During this time the early embryo is completely dependent on the oviduct and uterine environment for its survival and it is likely that the embryo requires an optimal uterine environment to ensure normal growth and viability. There is increasing evidence of an association between the concentration of systemic progesterone and early embryo loss and that progesterone supplementation of cows, particularly those with low progesterone, can reduce this loss. While progesterone is known to affect uterine function and embryo growth, little is known about the uterus during the period of early embryo loss and how this is affected by changes in the concentration of systemic progesterone. The expression of uterine genes encoding the transport protein retinol binding protein (RBP) and the gene for folate binding protein (FBP) appear to be sensitive to changes in systemic progesterone, particularly during the early luteal phase of the cycle. Uterine concentrations of proteins also seem to be regulated by stage of cycle; however, their relationship with the systemic concentration of progesterone is unclear. There is an urgent need to characterize the uterine environment from a functional perspective during the early part of the luteal phase of the cycle, particularly in the high-producing cow, in order to understand the factors contributing to early embryo loss and in order to devise strategies to minimize or reduce this loss.     

雷公藤多苷联合孕三烯酮对子宫内膜异位症卵巢功能及血清IL-6，TNF-α，hs-CRP，VEGF及APN的影响

目的:研究雷公藤多苷联合孕三烯酮对子宫内膜异位症卵巢功能及血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF)、超敏C反应蛋白(hs-CRP)、血管内皮生长因子(VEGF)、脂联素APN水平的影响。方法:选取2014年7月至2015年7月我院接诊的子宫内膜异位症患者80例并随机分为两组。对照组采用孕三烯酮进行治疗,观察组采用雷公藤多苷联合孕三烯酮进行治疗,观察两组患者治疗前后血清IL-6、TNF-α、hs-CRP、VEGF、APN、E2、FSH水平、TDS评分的变化并评价治疗疗效。结果:治疗后,观察组血清IL-6、TNF-α、hs-CRP、VEGF、E2、FSH水平及TDS评分均明显低于对照组(P0.05),血清APN水平及总有效率均明显高于对照组(P0.05)。结论:雷公藤多苷联合孕三烯酮能够有效改善子宫内膜异位症患者的卵巢功能,可能与其调节血清IL-6、TNF-α、hs-CRP、VEGF和APN水平有关。     

A morphological study of uterine alterations in mice due to exposure to cadmium

We investigated the morphologic and molecular effects of exposure to cadmium (Cd) for 30 and 60 days on the uteri of mice. We assessed uterine morphometric measurements, eosinophilia, mast cell numbers, endometrial apoptosis, proliferation and estrogen receptor alpha (ERα) immunoreactivity. We examined vaginal smears that reflected the hormonal alterations in the female reproductive tract. Because the female reproductive tract exhibits different morphology at each stage of the estrous cycle, we sacrificed all animals at estrus to make appropriate comparisons. Female BALB/c mice were exposed to 200 ppm Cd in their drinking water for either 30 or 60 days. Cd exposure caused significant decreases in endometrial thickness and number of glands in estrus phase uteri. The endometrial eosinophilia in the groups exposed to Cd also decreased compared to controls. Cd exposure increased the number of mast cells. Luminal and glandular epithelia were examined using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and by immunostaining proliferating cell nuclear antigen (PCNA) and estrogen receptor α (ERα). Compared to controls, the apoptotic index increased with time in both Cd exposed groups, while the proliferation index decreased. ERα immunoreactivity was decreased in both Cd exposed groups compared to controls; the decrease was most apparent in the 30 day Cd group. We found that 60 day Cd exposure increased apoptosis in the endometrium, which may affect the receptivity of the uterus for implantation.     

Current status of fertility control methods in India

Approximately 48.2% of couples of 15 to 49 years of age practice family planning methods in India. Female sterilization accounts for 34.2%, with male sterilization declining from 3·4% in 1992–93 to 1·9% in 1998–99. Use of the condom increased to 3·1% from 2·4%. There is an urgent need for research to develop new contraceptive modalities especially for men and also for women and to make existing methods more safe, affordable and acceptable. Current efforts in India to develop a male contraceptive are mainly directed towards (i) development of antispermatogenic agents to suppress sperm production, (ii) prevention of sperm maturation, (iii) prevention of sperm transport through vas deferens or rendering these sperm infertile and (iv) prevention of sperm deposition. Research work in the field of prevention of sperm transport through vas deferens has made significant advances. Styrene maleic anhydride (SMA) disturbed the electrical charge of spermatozoa leading to acrosome rupture and consequent loss in fertilizing ability of sperm. A multicentre phase-III clinical trial using SMA is continuing and it is hoped that the SMA approach would be available in the near future as an indigenously developed injectable intra-vasal male contraceptive. The safety and efficacy of available oral contraceptives were evaluated. An indigenously developed oral contraceptive ‘Centchorman’, which is a nonsteroidal, weakly estrogenic but potently antiestrogenic, was found to be safe and effective and is now being marketed in India since 1991 as a ‘once a week’ pill. Cyclofem and Mesigyna have been recommended as injectable contraceptives with proper counselling and service delivery by Indian studies. It has been recommended that these injectable contraceptives be added to the existing range of contraceptive methods available in the National Family Planning Programme. Based on the Indian studies CuT 200 was also recommended. Studies have indicated the advantage of intrauterine devices (IUD); they are long acting, relatively easily removed and fertility returns rapidly after their removal. Recent studies have recommended CuT 200 for use up to 5 years. The combination of some plant products i.e.Embelia ribes, Borax andPiper longum has been found to be safe and effective as a female contraceptive and the results of phase-I clinical trials are encouraging. Research work is going on in the country in various areas with special reference to hormonal contraceptive — a three monthly injectable contraceptive, immuno-contraceptives, antiprogestins, etc.