Reproductive Tract Epithelium Research Articles

Expression of Interferon Epsilon in Mucosal Epithelium is Regulated by Elf3.

Interferon epsilon (IFNε) is a unique type I interferon (IFN) that shows distinct constitutive expression in reproductive tract epithelium. Understanding how IFNε expression is regulated is critical for the mechanism of action in protecting the mucosa from infection. Combined computational and experimental investigation of the promoter of IFNε predicted transcription factor binding sites for the ETS family of transcription factors. We demonstrate here that Ifnε is regulated by Elf3, an epithelial restricted member of the ETS family. It is co-expressed with IFNε at the epithelium of uterus, lung and intestine, and we focused on regulation of IFNε expression in the uterus. Promoter reporter studies demonstrated that Elf3 was a strong driver of Ifnε expression; knockdown of Elf3 reduced expression levels of IFNε; Elf3 regulated Ifnε expression and chromatin immunoprecipitation (ChIP) confirmed the direct binding of Elf3 to the IFNε promoter. These data show that Elf3 is important in regulating protective mucosal immunity by driving constitutive expression of IFNε to protect mucosal tissues from infection in at least three organ systems.

Development and characterization of human fetal female reproductive tract organoids to understand Müllerian duct anomalies

Müllerian ducts are paired tubular structures that give rise to most of the female reproductive organs. Any abnormalities in the development and differentiation of these ducts lead to anatomical defects in the female reproductive tract organs categorized as Müllerian duct anomalies. Due to the limited access to fetal tissues, little is understood of human reproductive tract development and the associated anomalies. Although organoids represent a powerful model to decipher human development and disease, such organoids from fetal reproductive organs are not available. Here, we developed organoids from human fetal fallopian tubes and uteri and compared them with their adult counterparts. Our results demonstrate that human fetal reproductive tract epithelia do not express some of the typical markers of adult reproductive tract epithelia. Furthermore, fetal organoids are grossly, histologically, and proteomically different from adult organoids. While external supplementation of WNT ligands or activators in culture medium is an absolute requirement for the adult reproductive tract organoids, fetal organoids are able to grow in WNT-deficient conditions. We also developed decellularized tissue scaffolds from adult human fallopian tubes and uteri. Transplantation of fetal organoids onto these scaffolds led to the regeneration of the adult fallopian tube and uterine epithelia. Importantly, suppression of Wnt signaling, which is altered in patients with Müllerian duct anomalies, inhibits the regenerative ability of human fetal organoids and causes severe anatomical defects in the mouse reproductive tract. Thus, our fetal organoids represent an important platform to study the underlying basis of human female reproductive tract development and diseases.

Primary fallopian tube epithelial cells are resistant to Chlamydia trachomatis infection due to high expression of ido1.

Abstract Chlamydia trachomatis (Ct) is the most prevalent sexually transmitted bacterial infection in the world. In ~50% of cisgender women, Ct ascends from the cervix to infect the upper genital tract and increased ascension is linked to hormonal therapies. Oviduct infection induces inflammation which can lead to tubal scarring and sequelae of infertility and ectopic pregnancy. Fortunately, these complications occur in a minority of infected women, suggesting that intrinsic factors may protect the fallopian tubes from infection or tissue damage. In vitro inoculation of primary fallopian tube epithelial (FTE) cells with Ct serovar E resulted in a 10-fold reduction in infected cells compared to transformed endocervical HeLa cells. Indoleamine 2,3-dioxygenase (IDO) in the cytoplasm of epithelial cells acts as a key mediator of chlamydial host defense by metabolizing tryptophan, an essential Ct nutrient. We determined that uninfected primary FTE cells expressed on average 9 times higher levels of ido1 transcripts by qPCR compared to HeLa cells, and that in vitro infectivity of FTE cells was rescued by addition of an IDO-inhibitor (9-fold), tryptophan (21-fold), or indole (a metabolite that bypasses tryptophan starvation; 16-fold). Induction of IDO has mostly been attributed to IFNγ, but it can also be induced by Type I IFNs. Detection of high constitutive ido1 transcription in primary FTE cells indicates their ability to produce IDO independently of IFNγ. Ongoing studies are examining the role of Type I IFNs, including IFNɛ, which is constitutively expressed by reproductive tract epithelium and regulated by hormones. This work has implications for defining factors that contribute to protection of women from Ct-induced disease. The study was funded through a Translational Team Science Award (UNC School of Medicine and NC TRaCS) and also funded by NIAID NIH R01 AI067678 to U.M.N., NIAID NIH R01 AI119164 to T.D., and NIH grant DK065988 and Cystic Fibrosis Foundation grant BOUCHE15R0 to S.H.R.

Bi-allelic mutations in MCIDAS and CCNO cause human infertility associated with abnormal gamete transport.

Reduced generation of multiple motile cilia (RGMC) and the consequent primary ciliary dyskinesia (PCD) cause infertility due to a substantial reduction in the number of multiciliated cells (MCCs) in the efferent ducts (EDs)/oviducts. MCIDAS acts upstream of CCNO to regulate the biogenesis of basal bodies (BBs); therefore, both genes play a vital role in the multiciliogenesis of the reproductive tract epithelium. In this study, whole-exome sequencing was performed to identify the causative genes in 10 unrelated infertile patients with PCD: seven males and three females. Notably, homozygous frameshift mutations in MCIDAS (c.186dupT, p.Pro63Serfs*22) and CCNO (c.262_263insGGCCC, p.Gln88Argfs*8) were identified in one male and one female participant from two unrelated consanguineous families. Haematoxylin-eosin staining/scanning electron microscopy revealed abnormal MCCs in the mutated EDs/oviducts. Furthermore, transmission electron microscopy revealed significantly reduced BBs. Immunofluorescence staining showed the absence of MCIDAS and CCNO signals in the affected tissues and confirmed that MCIDAS acts upstream of CCNO in the context of multiciliogenesis in the reproductive tract epithelium. In vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) was successful, with a positive pregnancy outcome in both MCIDAS- and CCNO-mutated patients. Our results support the use of IVF/ICSI interventions to treat infertility due to RGMC in couples.

Using the Air-Liquid Interface Approach to Foster Apical-Basal Polarization of Mammalian Female Reproductive Tract Epithelia In Vitro.

Oviduct and uterus are key female reproductive organs lined by ciliated simple columnar epithelia, which are the first line of maternal contact with gametes and the developing embryo during reproduction and which warrant the optimal developmental environment for the conceptus. A major challenge for modeling these epithelia in vitro is the preservation of apical-basal polarization and cilia formation. The air-liquid interface (ALI) culture approach is a technology originally invented for modeling epidermal and airway epithelia. It has recently been shown that it also allows the establishment of highly differentiated in vitro models of epithelia that do not have access to ambient air in vivo. In this chapter, we present a comprehensive ALI procedure to model female reproductive tract (FRT) epithelia of different mammalian species in vitro over extended time periods. As a working example, the protocol focuses on primary oviductal epithelial cells (OEC) isolated from domestic pig. Hints on protocol variations for the culture of OEC from other species are provided in the Subheading 4.

Mini-Percoll Technique Induces Similar Capacitation Features in Domestic Ruminant Frozen-Thawed Spermatozoa Regardless of the Presence of Heparin

Background: Sperm capacitation is a process consists of a series of functional, biochemical, and biophysical modifications that render the ejaculated sperm competent for oocyte fertilization. Secreted by the female reproductive tract epithelium, heparin promotes capacitation by binding to and removing seminal plasma proteins, which are adsorbed to the sperm PM and would inhibit capacitation. There is substantial evidence that cryopreservation promotes capacitation-like changes in bull, ram and buck sperm. Our general hypotheses were: (a) cryopreserved ram sperm suffer capacitation more quickly than buck and bull sperm under the same conditions; (b) the capacitation status of ruminant cryopreserved sperm is similar whether or not heparin is present after the mini-Percoll technique; and (c) ruminant frozen-thawed sperm selected by mini-Percoll and incubated within media without heparin supplementation is not impaired in terms of capacitation status and sperm agglutination. This study aimed to compare sperm parameters of ovine, caprine, and bovine frozen-thawed sperm after mini-Percoll processing followed by incubation with or without heparin supplementation.Materials, Methods & Results: Commercial semen of all species were used. Sperm samples were selected by mini-Percoll and supplemented (or not) with heparin within an incubation medium for 18 h. Sperm kinematics (CASA system analyzes), capacitation status (CTC staining) and sperm agglutination were evaluated after thawing, mini-Percoll, 1.5 h, 3 h, 6 h and 18 h. In comparison with post-thawing analysis, ovine species demonstrated a reduction (P < 0.05) in most of the sperm motility parameters after mini-Percoll. Conversely, ovine samples presented the highest (P < 0.05) rate of acrosome-reacted cells after mini-Percoll. Heparin supplementation did not affect most of the parameters evaluated (P > 0.05). In caprine and bovine species, a lower (P < 0.05) rate of sperm agglutination was observed in the presence of heparin at 18 h of incubation. In the absence of heparin, ovine samples showed a higher (P < 0.05) agglutination rate compared to the bovine species after long incubation period.Discussion: The present study compared sperm parameters (sperm kinematics, agglutination rate and capacitation status) of ruminant frozen-thawed sperm after mini-Percoll selection followed by in vitro incubation with or without heparin supplementation. In this study, it was observed the same rate of capacitated cells after the sperm selection (min-Percoll) between ruminant species. This indicate that the capacitation process occurs similarly between ruminant species, refuting the first hypothesis of this study. The presence of heparin did not influence the capacitation status of ruminant frozen-thawed sperm after mini-Percoll selection, it demonstrates that the second hypothesis was supported by this study making more economic and practical the use of ruminant frozen-thawed semen. The absence of heparin in the incubation medium did not harmed the capacitation status and sperm agglutination of ruminant frozen-thawed sperm. This supported the third hypothesis of the current study and indicate that the use of mini-Percoll technique regardless the presence of heparin could be a useful alternative for the preparation of ruminant frozen-thawed sperm. In conclusion, the capacitation status of ruminant frozen-thawed sperm is similar whether or not heparin is present after the mini-Percoll technique.

Air-liquid interface cell culture: From airway epithelium to the female reproductive tract.

The air-liquid interface (ALI) approach is primarily used to mimic respiratory tract epithelia in vitro. It is also known to support excellent differentiation of 3D multilayered skin models. To establish an ALI culture, epithelial cells are seeded into compartmentalized culture systems on porous filter supports or gel substrata. After an initial propagation period, the culture medium is removed from the apical side of the epithelium, exposing the cells to the surrounding air. Therefore, nutritive supply to the cells is warranted only by the basolateral cell pole. Under these conditions, the epithelial cells differentiate and regain full baso-apical polarity. Some types of epithelia even generate in vivo-like apical fluid or mucus. Interestingly, the ALI culture approach has also been shown to support morphological and functional differentiation of epithelial cells that are not normally exposed to ambient air in vivo. This review aims at giving a brief overview on the characteristics of ALI cultures in general and ALI models of female reproductive tract epithelia in particular. We discuss the applicability of ALI models for the investigation of the early embryonic microenvironment and for its implications in assisted reproductive technologies.

Elongation of Müllerian ducts and connection to urogenital sinus determine the borderline of uterine and vaginal development

In female mice, proximal, middle and caudal Müllerian ducts (MDs) differentiate into oviduct, uterus and vagina, respectively. The fates of female reproductive tract epithelia are determined by the mesenchyme. However, the mesenchymal fate determination system is still unclear. It is reported that presence or absence of retinoic acid (RA) signaling in MD mesenchyme induced uterine or vaginal mesenchyme, respectively. To analyze determination of the borderline, RA signal switching factors were found to play critical roles. Expression of a RA metabolizing enzyme, CYP26A1, was high in the epithelium of caudal MD and urogenital sinus, indicating that the enzyme causes the absence of RA signaling in the region. mRNA expression of some transcription factors regulating Aldh1a2, RA synthesis enzyme expressed in MDs, in other tissues was detected in MDs. When the transcription factor genes were overexpressed in a uterine mesenchymal cell line, C/ebpδ overexpression stimulated Aldh1a2 expression. Furthermore, C/EBPδ protein was strongly expressed in the proximal and middle regions of the MDs and bound to the Aldh1a2 promoter in vivo. Since C/ebpδ mRNA expression was maintained at the same level in proximal, middle and caudal MDs, we hypothesize that a high frequency of mitosis induces a low level protein expression in MD mesenchyme. In fact, the mitotic activity was significantly high in caudal mesenchyme, and a mathematical model showed that a gradient of protein was induced by cell proliferation. Therefore, morphogenesis of MDs controls the fate of mesenchyme via RA degradation in urogenital sinus and a gradient of proteins involved in RA synthesis.

A morphological study of the male reproductive tract, post-testicular acrosome maturation and spermatophore formation in the black tiger prawn (Penaeus monodon).

Inferior larval production rates of domesticated Penaeus monodon broodstock has been a major hurdle to the expansion of its aquaculture, so that a better understanding of basic male reproductive biology is critical to improve the reproductive performance of this commercially important penaeid species. Following our previous study of spermatogenesis in the species, this study explored the mechanism of spermatophore formation with regards to the contribution of the reproductive tract epithelium by light and transmission electron microscopy. Four types of epithelial secretions (S1-4) were observed contributing to the three layers of the spermatophore. The primary layer of spermatophore was composed of S1 and S2, which were released from the secretory epithelial cells of the proximal vas deferens (PVD) and the secretory epithelial cells of the sperm-bearing lumen of the mid vas deferens, respectively. The secondary layer of the spermatophore was composed of S3, the secretory product of epithelial cells in the accessory tubule lumen of the mid vas deferens. The outer layer of the spermatophore was formed from S4 which was secreted by the epithelial cells in the posterior mid vas deferens and the terminal ampulla. Unique folds of the vas deferens epithelium appeared to play an important role in the formation of the spermatophore, particularly in the formation of the laminated structure of the spermatophore appendage. With respect to acrosome maturation along the reproductive tract, most spermatozoa did not have a fully developed anterior spike and a subacrosomal region when in the PVD, whereas both structures were fully formed by the time the spermatozoa reached the mid vas deferens and increased electron density when in the terminal ampulla; this observation represents the first morphological evidence of post-testicular acrosome maturation in this taxon.

Interferon-gamma inhibits seminal plasma induction of colony-stimulating factor 2 in mouse and human reproductive tract epithelial cells.

Seminal fluid interacts with the female reproductive tract to initiate a permissive immune response that facilitates embryo implantation and pregnancy success. The immune-regulatory cytokine interferon-γ (IFNG), which can be elevated in seminal plasma, is associated with reduced fertility. Here, we investigated how IFNG influences the female immune response to seminal fluid. In human Ect1 cervical epithelial cells, IFNG added at physiologically relevant concentrations substantially impaired seminal plasma-induced synthesis of key cytokines colony-stimulating factor 2 (CSF2) and interleukin-6 (IL6). Seminal fluid-induced CSF2 synthesis was also suppressed in the uterus of mice in vivo, when IFNG was delivered transcervically 12 h after mating. Transforming growth factor B1 (TGFB1) is the major seminal fluid signaling factor which elicits CSF2 induction, and IFNG exhibited potent dose-dependent suppression of CSF2 synthesis induced by TGFB1 in murine uterine epithelial cells in vitro. Similarly, IFNG suppressed TGFB1-mediated CSF2 induction in Ect1 cells and human primary cervical epithelial cells; however, IL6 regulation by IFNG was independent of TGFB1. Quantitative PCR confirmed that CSF2 regulation by IFNG in Ect1 cells occurs at the gene transcription level, secondary to IFNG suppression of TGFBR2 encoding TGFB receptor 2. Conversely, TGFB1 suppressed IFNG receptor 1 and 2 genes IFNGR1 and IFNGR2. These data identify IFNG as a potent inhibitor of the TGFB-mediated seminal fluid interaction with relevant reproductive tract epithelia in mice and human. These findings raise the prospect that IFNG in the male partner's seminal fluid impairs immune adaptation for pregnancy following coitus in women.

Modeling early embryo-maternal interactions in vitro

Environmental conditions experienced during early embryonic development influence growth, metabolism, and gene expression of the embryo as well as the epigenetic profile of the offspring. The environment of the early embryo consists of the luminal fluid within the oviduct and uterus and the epithelial cells composing this fluid. Whether the embryo is able to shape its own microenvironment by interacting with the epithelial lining of the oviduct/uterus and which factors potentially interfere with or regulate these interactions remains to be elucidated. As early embryonic signals and the respective maternal responses are subtle and local events, it is challenging to study them in vivo. Therefore, adequate in vitro-models optimally mimicking the contact zone between the maternal reproductive tract and the early embryo are needed to a) elucidate basic mechanisms involved in early embryonic development and b) reduce the number of experimental animals used for such studies. Functional epithelial cells are generally defined by a polarized distribution of organelles and proteins. Proper polarization is tightly connected with physiological cell behavior and in vivo-like reactivity of the epithelium. Therefore, this review summarizes current strategies for in vitro preservation of epithelial cell polarity. It presents recent advances in 3D culture of female reproductive tract epithelia and embryo-epithelial co-cultures. A special emphasis is set on compartmentalized culture systems, powerful tools for studying early embryo-maternal interactions in vitro. In such systems, cultured epithelial cells are manipulable from their basolateral as well as their apical cell pole, allowing concomitant application of embryonic as well as maternal effectors from the appropriate cellular compartment.

Mucosal Humoral Immune Response to SIVmac239∆nef Vaccination and Vaginal Challenge.

Live attenuated vaccines such as SIV with a deleted nef gene have provided the most robust protection against subsequent vaginal challenge with wild-type (WT) SIV in the SIV-rhesus macaque model of HIV-1 transmission to women. Hence, identifying correlates of this protection could enable design of an effective HIV-1 vaccine. One such prechallenge correlate of protection from vaginal challenge has recently been identified as a system with three components: 1) IgG Abs reacting with the viral envelope glycoprotein trimeric gp41; 2) produced by plasma cells in the submucosa and ectopic tertiary lymphoid follicles in the ectocervix and vagina; and 3) concentrated on the path of virus entry by the neonatal FcR in the overlying epithelium. We now examine the mucosal production of the Ab component of this system after vaginal challenge. We show that vaginal challenge immediately elicits striking increases in plasma cells not only in the female reproductive tract but also at other mucosal sites, and that these increases correlate with low but persistent replication at mucosal sites. We describe vaginal ectopic follicles that are structurally and functionally organized similar to follicles in secondary lymphoid organs, and we provide inferential evidence for a key role of the female reproductive tract epithelium in facilitating Ab production, affinity maturation, and class switch recombination. Vaccination thus accesses an epithelial-immune system axis in the female reproductive tract to respond to exposure to mucosal pathogens. Designing strategies to mimic this system could advance development of an effective HIV-1 vaccine.

Modeling the transcriptome of genital tract epithelial cells and macrophages in healthy mucosa versus mucosa inflamed by Chlamydia muridarum infection.

Chlamydia trachomatis urogenital serovars are intracellular bacteria that parasitize human reproductive tract epithelium. As the principal cell type supporting bacterial replication, epithelial cells are central to Chlamydia immunobiology initially as sentries and innate defenders, and subsequently as collaborators in adaptive immunity-mediated bacterial clearance. In asymptomatic individuals who do not seek medical care a decisive struggle between C. trachomatis and host defenses occurs at the epithelial interface. For this study, we modeled the immunobiology of epithelial cells and macrophages lining healthy genital mucosa and inflamed/infected mucosa during the transition from innate to adaptive immunity. Upper reproductive tract epithelial cell line responses were compared to bone marrow-derived macrophages utilizing gene expression microarray technology. Those comparisons showed minor differences in the intrinsic innate defenses of macrophages and epithelial cells. Major lineage-specific differences in immunobiology relate to epithelial collaboration with adaptive immunity including an epithelial requirement for inflammatory cytokines to express MHC class II molecules, and a paucity and imbalance between costimulatory and coinhibitory ligands on epithelial cells that potentially limits sterilizing immunity (replication termination) to Chlamydia-specific T cells activated with limited or unconventional second signals.

BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior.

Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success. In Drosophila melanogaster, these signals are generated by a variety of seminal peptides, many produced by the accessory glands (AGs). One epithelial cell type in the adult male AGs, the secondary cell (SC), grows selectively in response to bone morphogenetic protein (BMP) signaling. This signaling is involved in blocking the rapid remating of mated females, which contributes to the reproductive advantage of the first male to mate. In this paper, we show that SCs secrete exosomes, membrane-bound vesicles generated inside late endosomal multivesicular bodies (MVBs). After mating, exosomes fuse with sperm (as also seen in vitro for human prostate-derived exosomes and sperm) and interact with female reproductive tract epithelia. Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling. Indeed, when BMP signaling was reduced in SCs, vesicles were still formed in MVBs but not secreted as exosomes. These results demonstrate a new function for the MVB-exosome pathway in the reproductive tract that appears to be conserved across evolution.

An atypical CD8 T‐cell response to Chlamydia muridarum genital tract infections includes T cells that produce interleukin‐13

Chlamydia trachomatis urogenital serovars D-K are intracellular bacterial pathogens that replicate almost exclusively in human reproductive tract epithelium. In the C. muridarum mouse model for human Chlamydia genital tract infections CD4 T helper type 1 cell responses mediate protective immunity while CD8 T-cell responses have been associated with scarring and infertility. Scarring mediated by CD8 T cells requires production of tumour necrosis factor-α (TNF-α); however, TNF-α is associated with protective immunity mediated by CD4 T cells. The latter result suggests that TNF-α in-and-of itself may not be the sole determining factor in immunopathology. CD8 T cells mediating immunopathology presumably do something in addition to producing TNF-α that is detrimental during resolution of genital tract infections. To investigate the mechanism underlying CD8 immunopathology we attempted to isolate Chlamydia-specific CD8 T-cell clones from mice that self-cleared genital tract infections. They could not be derived with antigen-pulsed irradiated naive splenocytes; instead derivation required use of irradiated immune splenocyte antigen-presenting cells. The Chlamydia-specific CD8 T-cell clones had relatively low cell surface CD8 levels and the majority were not restricted by MHC class Ia molecules. They did not express Plac8, and had varying abilities to terminate Chlamydia replication in epithelial cells. Two of the five CD8 clones produced interleukin-13 (IL-13) in addition to IL-2, TNF-α, IL-10 and interferon-γ. IL-13-producing Chlamydia-specific CD8 T cells may contribute to immunopathology during C. muridarum genital tract infections based on known roles of TNF-α and IL-13 in scar formation.

Immunohistochemical assessment of epithelial integrity following intrauterine administration of polidocanol foam in Rheus macaques

Intrauterine administration of polidocanol foam causes tubal occlusion, providing a nonsurgical method of permanent contraception. Reliable tubal blockade requires multiple treatments, indicating that the oviduct has a robust ability to re-epithelialize after sclerosant therapy. The objective of this study was to characterize the reproductive tract epithelium after polidocanol treatment.

Manifestations of immune tolerance in the human female reproductive tract

Like other mucosal surfaces (e.g., the gastrointestinal tract, the respiratory tract), the human female reproductive tract acts as an initial barrier to foreign antigens. In this role, the epithelial surface and subepithelial immune cells must balance protection against pathogenic insults against harmful inflammatory reactions and acceptance of particular foreign antigens. Two common examples of these acceptable foreign antigens are the fetal allograft and human semen/sperm. Both are purposely deposited into the female genital tract and appropriate immunologic response to these non-self antigens is essential to the survival of the species. In light of the weight of this task, it is not surprising that multiple, redundant and overlapping mechanisms are involved. For instance, cells at the immunologic interface between self (female reproductive tract epithelium) and non-self (placental trophoblast cells or human sperm) express glycosylation patterns that mimic those on many metastatic cancer cells and successful pathogens. The cytokine/chemokine milieu at this interface is altered through endocrine and immunologic mechanisms to favor tolerance of non-self. The “foreign” cells themselves also play an integral role in their own immunologic acceptance, since sperm and placental trophoblast cells are unusual and unique in their antigen presenting molecule expression patterns. Here, we will discuss these and other mechanisms that allow the human female reproductive tract to perform this delicate and indispensible balancing act.

Generation of Human Female Reproductive Tract Epithelium from Human Embryonic Stem Cells

BackgroundRecent studies have identified stem/progenitor cells in human and mouse uterine epithelium, which are postulated to be responsible for tissue regeneration and proliferative disorders of human endometrium. These progenitor cells are thought to be derived from Müllerian duct (MD), the primordial female reproductive tract (FRT).Methodology/Principal FindingsWe have developed a model of human reproductive tract development in which inductive neonatal mouse uterine mesenchyme (nMUM) is recombined with green fluorescent protein (GFP)-tagged human embryonic stem cells (hESCs); GFP-hESC (ENVY). We demonstrate for the first time that hESCs can be differentiated into cells with a human FRT epithelial cell phenotype. hESC derived FRT epithelial cells emerged from cultures containing MIXL1+ mesendodermal precursors, paralleling events occurring during normal organogenesis. Following transplantation, nMUM treated embryoid bodies (EBs) generated epithelial structures with a typical MD phenotype that expressed the MD markers PAX2, HOXA10. Functionally, the hESCs derived FRT epithelium responded to exogenous estrogen by proliferating and secreting uterine-specific glycodelin A (GdA).Conclusions/SignificanceThese data show nMUM can induce differentiation of hESC to form the FRT epithelium. This may provide a model to study early developmental events of the human FRT.

Molecular characterization of Lingual antimicrobial peptide in the female reproductive tract of Buffalo

Bubalus bubalis (Ruminantia: Bovidae, Bovinae) is an economically important animal of many Asian countries, making significant contribution to milk and meat production. Sub clinical infection of the reproductive tract is one of the important causes for reduced reproductive efficiency in dairy herd of buffaloes. Antimicrobial peptides are component of innate immune system which helps in augmenting the resistance to infection at epithelial surfaces e.g reproductive tract epithelium. In this study we have identified a s-defensin called Lingual Antimicrobial Peptide (LAP) in buffalo reproductive tract. Interestingly the gene was 100 % identical to the LAP isolated from the tongue epithelium of Bos taurus. The 195 bp cDNA of LAP codes for 64 amino acids and of which 50% are cationic amino acids. Phylogenetic studies indicate that LAP of reproductive epithelium of buffalo is different from other beta defensins isolated from the various tissues of same species, but all beta defensin were found to have the same progenitor gene. It is concluded that buffalo reproductive tract epithelium lining contains LAP.

Mǔllerian cyst of the mesentery: A case report of an unusual location

Mǔllerian cysts or paramesonephric cysts arise from the fused embryonic ducts, which typically regress in the uterus. These cysts are usually located paravertebrally. We present an unusual case of a Mǔllerian cyst developing within the mesentery of the ileocecum that was successfully resected. The patient presented to our surgical unit with abdominal pain and swelling. She underwent all the necessary tests to rule out other pathologies before she underwent right hemicolectomy. The patient was discharged without complications. Histopathology confirmed the presence of female reproductive tract epithelium, which was conclusive of a Mǔllerian cyst or paramesonephric cyst. Mǔllerian cysts are rarely malignant, and they are usually treated surgically. The incidence of Mǔllerian cysts is one in 105,000, with almost equal sex distribution. Their unusual intraperitoneal location further demonstrates their uncommon presentation.