Biobanking human endometrial tissue and blood specimens: standard operating procedure and importance to reproductive biology research and diagnostic development

Sheldon, Elizabeth; Vo, Kim Chi; McIntire, Ramsey; Aghajanova, Lusine; Zelenko, Zara; Irwin, Juan C.; Giudice, Linda C.

doi:10.1016/j.fertnstert.2011.01.164

Cited by 30 publications

(30 citation statements)

References 21 publications

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“…Only eutopic endometrial samples were studied. All patients undergoing surgery for endometriosis-related pain and/or infertility gave written informed consent through the UCSF NIH Human Endometrial Tissue and DNA Bank [18] (Supplemental Table S1; Supplemental Data are available online at www.biolreprod.org). Seventeen eutopic endometrial tissue samples were from patients with severe endometriosis (stage IV; n = 4 in proliferative [PE], n = 7 in early secretory [ESE], and n = 6 in midsecretory [MSE] phases).…”

Section: Methodsmentioning

confidence: 99%

Aberrant Endometrial DNA Methylome and Associated Gene Expression in Women with Endometriosis

Houshdaran

Nezhat

et al. 2016

Biology of Reproduction

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Endometriosis is an estrogen-dependent, progesterone-resistant disorder largely derived from retrograde transplantation of menstrual tissue/cells into the pelvis, eliciting an inflammatory response, pelvic pain, and infertility. Eutopic endometrium (within the uterus), giving rise to pelvic disease, displays cycle-dependent transcriptomic, proteomic, and signaling abnormalities, and although its DNA methylation profiles dynamically change across the cycle in healthy women, studies in endometriosis are limited. Herein, we investigated the DNA methylome and associated gene expression in three phases of the cycle in eutopic endometrium of women with severe endometriosis versus controls, matched for ethnicity, medications, smoking, and no recent contraceptive steroid use. Genome-wide DNA methylation and gene expression were coassessed in each sample. Cycle phase was determined by histology, serum hormone levels, and unsupervised principal component and hierarchical cluster analyses of microarray data. Altered endometrial DNA methylation in endometriosis was most prominent in the midsecretory phase (peak progesterone), with disruption of the normal pattern of cycle-dependent DNA methylation changes, including a bias toward methylation of CpG islands, suggesting wide-range abnormalities of the chromatin remodeling machinery in endometriosis. DNA methylation changes were associated with altered gene expression relevant to endometrial function/dysfunction, including cell proliferation, inflammation/immune response, angiogenesis, and steroid hormone response. The data provide insight into epigenetic reprogramming and steroid hormone actions in endometrium contributing to the pathogenesis and pathophysiology of endometriosis.

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Section: Methodsmentioning

confidence: 99%

Aberrant Endometrial DNA Methylome and Associated Gene Expression in Women with Endometriosis

Houshdaran

Nezhat

et al. 2016

Biology of Reproduction

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“…Deidentified decidual specimens from elective terminations (gestational age, 6.0 to 14.3 weeks) were collected. Deidentified endometrial specimens from benign hysterectomies were freshly acquired from the UCSF-NIH Human Endometrial Tissue and DNA Bank (64). Briefly (see reference 65 for the complete protocol), 2-mm 3 fragments of decidua parietalis or endometrium were microdissected; placed onto Transwell filters (30-mm diameter, 0.4 m, hydrophilic polytetrafluoroethylene [PTFE] membrane; Millipore) precoated with 0.1 ml Matrigel (BD Biosciences); and immediately cultured in Dulbecco modified Eagle medium (DMEM)-F-12 medium (1:1, vol/vol) with Gibco GlutaMAX (Thermo Fisher Scientific) supplemented with 2.5% fetal bovine serum, 100 IU/ml penicillin, 100 g/ml streptomycin, 50 g/ml gentamicin, and 1.25 g/ml amphotericin B (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

Limited Colonization Undermined by Inadequate Early Immune Responses Defines the Dynamics of Decidual Listeriosis

et al. 2017

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The bacterial pathogen Listeria monocytogenes causes foodborne systemic disease in pregnant women, which can lead to preterm labor, stillbirth, or severe neonatal disease. Colonization of the maternal decidua appears to be an initial step in the maternal component of the disease as well as bacterial transmission to the placenta and fetus. Host-pathogen interactions in the decidua during this early stage of infection remain poorly understood. Here, we assessed the dynamics of L. monocytogenes infection in primary human decidual organ cultures and in the murine decidua in vivo. A high inoculum was necessary to infect both human and mouse deciduas, and the data support the existence of a barrier to initial colonization of the murine decidua. If successful, however, colonization in both species was followed by significant bacterial expansion associated with an inability of the decidua to mount appropriate innate cellular immune responses. The innate immune deficits included the failure of bacterial foci to attract macrophages and NK cells, cell types known to be important for early defenses against L. monocytogenes in the spleen, as well as a decrease in the tissue density of inflammatory Ly6C hi monocytes in vivo. These results suggest that the infectivity of the decidua is not the result of an enhanced recruitment of L. monocytogenes to the gestational uterus but rather is due to compromised local innate cellular immune responses.KEYWORDS Listeria monocytogenes, decidua, placenta T he pregnant uterus is a dynamic organ that supports fetoplacental growth, with many trophic functions provided by the decidua, the specialized endometrial tissue that encases the conceptus (1). The decidua also provides a unique immunological environment that blocks the maternal immune system from recognizing the conceptus as a foreign tissue and rejecting it as if it were a solid-organ transplant (2). As a consequence, this environment would be expected to provide opportunities for pathogens to infect the maternal-fetal interface. Indeed, infection of the decidua is now considered to be a critical first step in the hematogenous transmission of a variety of pathogens from mother to fetus (3-7). One such organism is the facultative, intracellular, Gram-positive bacterium Listeria monocytogenes, a clinically relevant foodborne pathogen that can cause severe disease in pregnant women. Once L. monocytogenes reaches the placenta, there are increased risks of severe maternal disease and pregnancy complications, including preterm labor and devastating fetal/neonatal morbidity and mortality (8, 9).Because of its experimental tractability, L. monocytogenes is an excellent model

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“…Additional samples for validation studies (n = 3) were obtained in the proliferative phase. Tissue samples were obtained through the National Institutes of Health Specialized Cooperative Centers Program in Reproduction and Infertility Research Human Endometrial Tissue and DNA Bank at UCSF under established standard operating procedures (24). Endometrial tissue samples included six biopsies (obtained using the Pipelle Endometrial Suction Curette, Cooper Surgical) from subjects undergoing oocyte retrieval, hysteroscopy, or laparoscopic surgery for benign conditions and one hysterectomy specimen.…”

Section: Methodsmentioning

confidence: 99%

Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production

Chen

Erikson

Piltonen

et al. 2013

Fertility and Sterility

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Objective To determine the effects of coculturing endometrial epithelial cells (eEC) with paired endometrial stromal fibroblasts (eSF) on cell-specific gene expression and cytokine secretion patterns. Design In vitro study. Setting University research laboratory. Patient(s) Endometrial biopsies were obtained from premenopausal women. Intervention(s) Polarized eEC and subject-paired eSF were cultured for 12.5 hours alone (monoculture) or combined in a two-chamber coculture system without cell-cell contact. Cells and conditioned media were analyzed for global gene expression and cytokine secretion, respectively. Purified, endometrial tissue-derived eEC and eSF isolated by fluorescent activated cell sorting (FACS) were used as noncultured controls. Main Outcome Measure(s) Cell-specific global gene expression profiling and analysis of secreted cytokines in eEC/eSF cocultures and respective monocultures. Result(s) Transepithelial resistance, diffusible tracer exclusion, expression of tight junction proteins, and apical/basolateral vectorial secretion confirmed eEC structural and functional polarization. Distinct transcriptomes of eEC and eSF were consistent with their respective lineages and their endometrial origin. Coculture of eEC with eSF resulted in altered cell-specific gene expression and cytokine secretion. Conclusion(s) This coculture model provides evidence that interactions between endometrial functionally polarized epithelium and stromal fibroblasts affect cell-specific gene expression and cytokine secretion underscoring their relevance when modeling endometrium in vitro.

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Biobanking human endometrial tissue and blood specimens: standard operating procedure and importance to reproductive biology research and diagnostic development

Cited by 30 publications

References 21 publications

Aberrant Endometrial DNA Methylome and Associated Gene Expression in Women with Endometriosis

Aberrant Endometrial DNA Methylome and Associated Gene Expression in Women with Endometriosis

Limited Colonization Undermined by Inadequate Early Immune Responses Defines the Dynamics of Decidual Listeriosis

Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production

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