Sandra Anderson
Eastern Virginia Medical School
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Featured researches published by Sandra Anderson.
Molecular Human Reproduction | 2012
Hai Wang; Federica Pilla; Sandra Anderson; Sebastián Martínez-Escribano; Isabel Herrer; Juan Manuel Moreno-Moya; Sirisha Musti; Silvina Bocca; Sergio Oehninger; J.A. Horcajadas
There is an urgent need to develop optimized experimental models to examine human implantation. These studies aimed to (i) establish a human endometrium-like three-dimensional (3D) culture system, and (ii) examine the attachment of trophoblast-like Jar spheroids to the culture. In the present work, 3D endometrial cultures were constructed with fibrin-agarose as matrix scaffold, and using epithelial and stromal cells from both human primary cultures and established cell lines. An attachment assay between trophoblast cells and the 3D culture was developed. Epithelial cells (cytokeratin(+)) concentrated on top of the matrix forming a monolayer, and stromal cells (vimentin(+)) resided within the matrix, resembling the normal endometrial structure. The capability of primary epithelial cells to form glands spontaneously was observed. Human trophoblast cells (Jar cells) were hCG(+) by immunostaining, allowed to form spheroids, and confirmed to secrete hCG into the medium. Time-dependent experiments demonstrated a high rate of attachment of Jar spheroids to the epithelium, and adhesion was strongly related to the various cell types present in the 3D culture. An architecturally and functionally competent 3D endometrial culture system was established, that coupled with Jar spheroids mimicking trophoblast cells, provides a unique in vitro model for the study of certain aspects of human implantation.
Molecular Human Reproduction | 2011
A. Franchi; Silvina Bocca; Sandra Anderson; Ryan M. Riggs; Sergio Oehninger
Milk fat globule epidermal growth-factor 8 (MFG-E8) has not been previously linked to endometrial physiology. We reported on MFG-E8 mRNA up-regulation in the human endometrium during the window of implantation (WOI) using microarrays. Prolactin (PRL) secreted by stromal cells has been suggested to modulate protein expression. The objective of this study was to characterize the endometrial expression of MFG-E8 and its ligand αvβ3 integrin during the menstrual cycle and its possible regulation by PRL. MFG-E8 mRNA (real-time RT-PCR) and protein expression (immunohistochemistry and immunoblotting) were analyzed in human endometrial biopsies at different times of the menstrual cycle, as well as in primary endometrial cell cultures. In primary cultures of epithelial cells, MFG-E8 intracellular protein expression was evaluated in absence or presence of PRL (0.2 and 1 μg/ml). The results show that MFG-E8 protein is almost exclusively localized to the epithelium in whole endometrial biopsies. Both MFG-E8 mRNA and protein expression increased in the luteal phase and were highest during the WOI; epithelial protein location of αvβ3 integrin also peaked on cycle Day 24. Cultured epithelial cells showed a diffuse staining of MFG-E8 over the cytoplasmic area; however, some cells presented a punctuated staining pattern. PRL treatment of epithelial cells for 72 h in vitro significantly increased MFG-E8 protein intracellular expression. This is the first report on MFG-E8 protein localization to the human endometrial epithelium and its up-regulation during the WOI. The pattern of glandular expression of its ligand αvβ3 integrin was remarkably similar. In vitro data support a modulatory role for PRL as a stromal/epithelial paracrine factor controlling MFG-E8.
Placenta | 2012
Silvina Bocca; Sandra Anderson; B. Amaker; R.J. Swanson; A. Franchi; F. Lattanzio; Sergio Oehninger
OBJECTIVES MFG-E8 is a novel endometrial protein with conserved functions in tissue remodeling and angiogenesis in non-uterine tissues. Our aims were: 1. To examine the presence of MFG-E8 protein in the human endometrium during the window of implantation, in human endometrial cell lines, in human placental tissue at different gestational ages, and in murine implantation sites during early gestation; and 2. To study the regulation of MFG-E8 mRNA expression in mice implantation sites. STUDY DESIGN MFG-E8 protein and its receptor integrin αvβ3 were detected by immunostaining in human endometrial biopsies obtained from normal volunteers, in human endometrial cell lines (epithelial: Ishikawa and HEC-1A, stromal: HESC, and endothelial: HEEC), in human products of conception from all trimesters of gestation, and in murine implantation and inter-implantation sites dissected on days 5 and 8 post-coitus. MFG-E8 gene expression was assessed by RT-PCR. MAIN OUTCOME MEASURES Immunohistochemical determination of MFG-E8 in endometrium and products of conception as well as relative MFG-E8 mRNA expression in mice implantation sites. RESULTS MFG-E8 protein was present almost exclusively in the epithelial compartment of human endometrium. It was also expressed in the cytotrophoblasts and syncytiotrophoblasts outlining chorionic villi of the human placenta at all trimesters of gestation, and in murine implantation sites. MFG-E8 mRNA was significantly up-regulated in murine implantation sites and with increased gestational age. CONCLUSIONS MFG-E8 expression in the endometrial epithelium as well as in chorionic villi suggests its possible role in endometrial reorganization during the receptive phase and in events related to normal pregnancy in mammals.
Fertility and Sterility | 2014
Carla Schmitz; Liang Yu; Silvina Bocca; Sandra Anderson; João Sabino Cunha-Filho; Sergio Oehninger
OBJECTIVE To investigate the role of MFG-E8 and its receptor integrin αvβ3 in the attachment of trophoblast cells to the endometrial epithelium. DESIGN Experimental in vitro study. SETTING Academic center. PATIENT(S) None. INTERVENTION(S) By using a well-differentiated endometrial adenocarcinoma cell line (Ishikawa cells) and choriocarcinoma human trophoblast cells (Jar cells), an in vitro assay mimicking human implantation was established. To investigate the impact of blocking MFG-E8 and integrin αvβ3, we pretreated the cell lines with antibodies against those proteins at different concentrations before the attachment assay. MAIN OUTCOME MEASURE(S) Attachment rate of Jar spheroids to the epithelial cell monolayer. RESULT(S) Pretreatment of Ishikawa cells with anti-MFG-E8 antibody caused a dose-dependent and significant inhibition of attachment. On the other hand, pretreatment of Jar spheroids did not result in a significant effect on the attachment rate. Pretreatment of Ishikawa cells as well as Jar spheroids with anti-integrin αvβ3 antibodies resulted in a dose-dependent, significant inhibition of attachment. CONCLUSION(S) This study showed that blocking MFG-E8 and its receptor integrin αvβ3 in Ishikawa cells diminishes Jar spheroid attachment. Moreover, blocking integrin αvβ3 in the trophoblastic cells also diminished their attachment to the Ishikawa monolayer.
Fertility and Sterility | 2014
Liang Yu; Sandra Anderson; Sergio Oehninger; Silvina Bocca
OBJECTIVE To explore the role of tumor necrosis factor (TNF) α, an early embryonic product, on endometrial epithelial cell migration and endometrial milk fat globule-epidermal growth factor 8 protein (MFG-E8) production. DESIGN In vitro study. SETTING Academic center. INTERVENTION(S) Ishikawa cells, used as surrogates for human epithelial cells, were treated with and without TNF-α. MAIN OUTCOME MEASURE(S) Effect of TNF-α on intracellular MFG-E8 protein was evaluated with the use of ELISA, Western blot, and subcellular fractionation. Specific inhibitors were used to study TNF-α mechanism of action. Effect of TNF-α on cell migration was studied with the use of a wound healing assay and reorganization of E-cadherin. RESULT(S) TNF-α induced: 1) significant up-regulation of MFG-E8 intracellular protein, which was attenuated by pretreatment with a specific inhibitor of nuclear factor κB; 2) increased transcription of MFG-E8 and other proinflammatory factors, such as interleukins 6 and 8, which were suppressed by cotreatment with hCG; and 3) significant cell migration with E-cadherin remodeling, changes associated with subcellular MFG-E8 relocalization. CONCLUSION(S) TNF-α up-regulates endometrial epithelial cell migration and MFG-E8 production, which are critical steps required for the endometrial changes during menstrual cycle as well as during embryonic attachment and invasion.
Cell signalling and Trafficking | 2013
Abbaa Sarhan; Silvina Bocca; Liang Yu; Sandra Anderson; Terry Jacot; Tanya C. Burch; Julius O. Nyalwidhe; Claretta Sullivan; Mandeep Kaur; Vladimir B. Bajic; Sergio Oehninger
Abstract Objective: We have recently showed that MFGE8, a novel epithelial cell protein in the human endometrium, upregulated during the window of implantation. We hypothesized that
Fertility and Sterility | 2012
Ryan M. Riggs; Silvina Bocca; Sandra Anderson; A. Franchi; Sergio Oehninger
OBJECTIVE To study the regulation of apoptosis in human endometrial cells. The specific aims were to determine whether milk fat globule-epidermal growth factor 8 (MFG-E8), a novel endometrial epithelial protein, modulates caspase activation and DNA fragmentation; and to examine whether hCG, an early embryonic product, regulates Bax and Bcl-2 equilibrium, as well as MFG-E8 expression. DESIGN Primary cultures of human endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs). SETTING Academic center. PATIENT(S) Ovulatory women aged 21-30 years. INTERVENTION(S) Treatment with MFG-E8 and hCG. MAIN OUTCOME MEASURE(S) Apoptotic activity was quantified using a luciferase assay. Deoxyribonucleic acid fragmentation was detected by TUNEL assay. Bax, Bcl-2, and MFG-E8 messenger RNA expression levels were determined by quantitative reverse transcription-polymerase chain reaction. Immunocytochemistry was used to establish cell purity and presence of MFG-E8 and hCG-R (receptor) proteins. RESULT(S) Endometrial epithelial cells were cytokeratin(+), vimentin(-), MFG-E8(+), and hCG-R(+), whereas ESC were vimentin(+), cytokeratin(-), MFG-E8(-), and hCG-R(+). Treatment of ESC with MFG-E8 resulted in a 13-fold increase in caspase activity and a 30-fold increase in TUNEL. On the other hand, hCG decreased messenger RNA expression of Bax in ESC. CONCLUSION(S) Milk fat globule-epidermal growth factor 8 has proapoptotic activity, suggesting participation in endometrial remodeling via an epithelial-stromal cell paracrine effect. Conversely, pregnancy levels of hCG has opposite effects on stromal cells.
Tissue Engineering Part C-methods | 2013
Hai Wang; Silvina Bocca; Sandra Anderson; Liang Yu; J.A. Horcajadas; Sergio Oehninger
Molecular Human Reproduction | 2010
Xiaomei Zhang; Silvina Bocca; A. Franchi; Sandra Anderson; Mandeep Kaur; Vladimir B. Bajic; Sergio Oehninger
Fertility and Sterility | 2010
Silvina Bocca; Sandra Anderson; Sergio Oehninger