Mitogenic actions of endothelin and other growth factors in ovine endometrium

1997 ◽  
Vol 152 (2) ◽  
pp. 283-290 ◽  
Author(s):  
L A Salamonsen ◽  
R J Young ◽  
S Garcia ◽  
J K Findlay
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Epithelial Cells ◽  
Stromal Cells ◽  
Cellular Proliferation ◽  
Calf Serum ◽  
Dose Dependence ◽  
Leukaemia Inhibitory Factor ◽  
Serum Free

Abstract Endothelin-1 (ET-1) is present in ovine endometrium, primarily in epithelial cells, and increases around the time of implantation. We examined the cell type expressing ET-binding sites in vitro and whether ET-1 has mitogenic actions in the endometrium, alone or in synergy with other growth factors. Purified epithelial and stromal cells were prepared from luteal-phase endometrium. Specific receptors were demonstrated by binding of 125I-ET-1 and proliferative effects of ET-1 and/or other growth factors determined by uptake of [3H]thymidine by cells in serum-free culture. 125I-ET-1 bound to both epithelial (2516 ± 820 c.p.m./well) and stromal (6368 ± 1350 c.p.m./well) cells and was displaced by ET-1 (1 μmol l−1). There were no proliferative effects of ET on epithelial cells. ET-1 (10 nmol l−1) stimulated uptake of [3H]thymidine by stromal cells under serum-free conditions in 13/20 individual cell preparations, to 149 ± 13% of control (untreated=100%) with dose-dependence between the range of 1 to 100 nmol l−1. Stimulation by fetal calf serum was to 377 ± 126% of control. The effects on proliferation by other growth factors (dose; % of control ± s.e.m., number of positives/total number of cell preparations) were: IGF-I (13 nmol l−1; 182 ± 14, 4/4), epidermal growth factor (EGF; 4·8 nmol l−1; 132 ± 5%, 7/7), platelet-derived growth factor-BB (0·4 nmol l−1; 146 ± 3, 2/2) and leukaemia inhibitory factor (0·4 nmol l−1; 110 ± 2, 3/3). All stimulations except that of EGF were significant and dose-responsive but only insulin was additive with ET (350 ± 35, 5/5). ET-1 also stimulated expression of the the AP-1 cis element c-jun, this being maximal at 60 min of exposure to mitogen. ET-1, along with other growth factors has a likely paracrine role in cellular proliferation in the endometrium, possibly in association with blastocyst implantation. Journal of Endocrinology (1997) 152, 283–290

229. Identification of transforming growth factor beta2 (TGF-β2) and its receptors TGF-βRI and TGF-βRII in the possum (Trichosurus vulpecula) prostate: evidence of seasonal changes

2008 ◽  
Vol 20 (9) ◽  
pp. 29
Author(s):  
H. Martyn ◽  
K. Pugazhenthi ◽  
B. McLeod ◽  
H. D. Nicholson
Keyword(s):  
Growth Factor ◽  
Epithelial Cells ◽  
Stromal Cells ◽  
Seasonal Changes ◽  
Transforming Growth Factor ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Western Blots ◽  
Prostate Growth

Benign Prostatic Hyperplasia is an enlargement of the prostate affecting the ageing male population. The common Brushtail possum (Trichosurus vulpecula) has been identified as a possible model to study factors regulating prostate growth because its prostate grows and regresses seasonally. Transforming growth factor Beta 2 (TGF-β2) is present in human prostatic tissue. In vitro, TGF-β inhibits epithelial cell, but stimulates stromal cell proliferation (Mori et al. 1990). TGF-β2 binds to TGF-β receptor II (TGF-βRII), which then recruits the type 1 receptor (TGF-βRI) (Saez et al. 1998) The aim of this study was to identify any seasonal changes in expression of TGF-β2 and its receptors in the possum prostate. Six wild-caught possums were sacrificed in each of the months of January, March, May, July, September and November. The prostates were divided into a cranial and caudal region and immunohistochemistry and Western Blot analysis performed. In each animal the glandular and periurethral areas of the caudal and cranial prostates were examined separately. Immunohistochemistry identified the presence of TGF-β2 in both the stromal and epithelial cells of the glandular and periurethral areas of the cranial and caudal regions. In the cranial tissue, more immuno-positive stromal cells than epithelial cells were present, whereas in the caudal tissue immuno-reactivity was predominantly localised to the epithelial cells. Analysis of the western blots suggested that TGF-β2 expression was lowest immediately before and during the breeding season (March, May). Both TGF-βRI and TGF-βRII were identified in all regions of the prostate. Furthermore, immunohistochemistry revealed that the receptors were co-localised in the epithelial and stromal cells in all areas. TGF-β2 and its receptors are present in the possum prostate. TGF-β2 localisation varies between the caudal and cranial regions and as predicted from in vitro experiments TGF-β2 expression decreases during prostate growth. (1) Mori H. et al. (1990). The Prostate, 16, 71 - 80. (2) Saez C. et al. (1998). The Prostate, 37, 84 - 90.

P–366 MicroRNAs at the embryo-maternal interface have effects on endometrial cell proliferation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Makri ◽  
M Castellanos-Uribe ◽  
S May ◽  
W Maalouf
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Growth Factors ◽  
Stromal Cells ◽  
Cellular Proliferation ◽  
Fold Change ◽  
Implantation Site ◽  
Endometrial Stromal Cells ◽  
Endometrial Cells

Abstract Study question Whether cell-free microRNAs are part of the embryo-maternal interactome with possible effects on processes related to implantation. Summary answer Specific microRNAs cause major transcriptomic changes in uterine cells and alter cellular proliferation which is pivotal for the implantation of the incoming embryo. What is known already A plethora of molecules present at the uterine luminal fluid including cytokines, growth factors, and adhesion proteins are involved in implantation. However little is known about the roles of extracellular microRNAs (miRNAs) at the embryo-maternal interface. MicroRNAs act mainly as gene regulators and a single miRNA can have thousands of gene targets. MiRNAs are released by blastocysts and uterine cells internalize miRNAs that are present in the extracellular environment. To date there is limited evidence on the molecular actions of these cell-free miRNAs and their effects on processes related to implantation. Study design, size, duration Human endometrial stromal cells (hESCs) were cultured in complete growth medium for 8 consecutive passages. A miRNA mimic experiment in 6 replications was carried out in which endometrial cells were transfected with miR–371a. Gene changes in the hESCs were studied with genome-wide microarray technology and the results were validated in vitro with PCR. Participants/materials, setting, methods The miR–371a mimic was transfected in hESCs using a Lipofectamine reagent. RNA was extracted and the samples were processed with microarray Clariom™ Human Assays using Affymetrix®. The transcriptomic profiles between transfected and control cells were compared using Partek®. Differentially expressed genes were considered significant when p-value was <0.05, false discovery rate, FDR ≤ 0.05 with Benjamini-Hochberg correction, and fold-change of > 1.5 or < –1.5. Functional enrichment analysis was carried out using WebGestalt and Enrichr. Main results and the role of chance MiR–371a altered the expression of 4.760 genes in endometrial cells (p < 0.05, fold-change 1.5). A total of 16 biological processes, 23 cellular components, and 24 molecular pathways were disrupted by this miRNA. WebGestalt analysis found 159 enriched categories including increase of negative cell cycle regulation, apoptosis signalling, and cycle arrest and decreased cell proliferation. Cell cycle was one of the most affected pathways in KEGG analysis with at least 54 genes dysregulated. Mammalian phenotype ontology analysis found 4.818 affected phenotypes, including decreased cell proliferation (58 genes), increased apoptosis (48 genes) and abnormal cell cycle (41 genes). Key-genes of endometrial proliferation at the window of implantation were significantly downregulated, including: CD44, PGR; IGFs, FGFs, and HAND2. Moreover, at least 25% decreased hESCs proliferation was verified in vitro after transfection. These negative effects of miR–371a in cell cycle could disturb implantation of the incoming embryo, since intense cellular proliferation is necessary for establishment of the implantation site. Limitations, reasons for caution These results are limited to miR–371a actions on human endometrial stromal cells. It is likely that miRNAs, cytokines, growth factors, and other molecules form complex regulatory networks that control uterine receptivity and embryo implantation. Wider implications of the findings: MiRNAs are important mediators of the embryo-maternal interactome. Their actions are likely involved in implantation-related processes including inter-cellular communication, decidualization, adhesion, invasion, and establishment of the implantation site. Embryo-secreted miRNAs change the transcriptome of the neighboring endometrial cells with effects on implantation-related pathways, serving thus secretory functions. Trial registration number N/A

scholarly journals IL-33 activates tumor stroma to promote intestinal polyposis

2015 ◽  
Vol 112 (19) ◽  
pp. E2487-E2496 ◽  
Author(s):  
Rebecca L. Maywald ◽  
Stephanie K. Doerner ◽  
Luca Pastorelli ◽  
Carlo De Salvo ◽  
Susan M. Benton ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Mast Cell ◽  
Growth Factors ◽  
Epithelial Cells ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Cell Activation ◽  
Intestinal Polyposis ◽  
Extracellular Matrix Components

Tumor epithelial cells develop within a microenvironment consisting of extracellular matrix, growth factors, and cytokines produced by nonepithelial stromal cells. In response to paracrine signals from tumor epithelia, stromal cells modify the microenvironment to promote tumor growth and metastasis. Here, we identify interleukin 33 (IL-33) as a regulator of tumor stromal cell activation and mediator of intestinal polyposis. In human colorectal cancer, IL-33 expression was induced in the tumor epithelium of adenomas and carcinomas, and expression of the IL-33 receptor, IL1RL1 (also referred to as IL1-R4 or ST2), localized predominantly to the stroma of adenoma and both the stroma and epithelium of carcinoma. Genetic and antibody abrogation of responsiveness to IL-33 in the ApcMin/+ mouse model of intestinal tumorigenesis inhibited proliferation, induced apoptosis, and suppressed angiogenesis in adenomatous polyps, which reduced both tumor number and size. Similar to human adenomas, IL-33 expression localized to tumor epithelial cells and expression of IL1RL1 associated with two stromal cell types, subepithelial myofibroblasts and mast cells, in ApcMin/+ polyps. In vitro, IL-33 stimulation of human subepithelial myofibroblasts induced the expression of extracellular matrix components and growth factors associated with intestinal tumor progression. IL-33 deficiency reduced mast cell accumulation in ApcMin/+ polyps and suppressed the expression of mast cell-derived proteases and cytokines known to promote polyposis. Based on these findings, we propose that IL-33 derived from the tumor epithelium promotes polyposis through the coordinated activation of stromal cells and the formation of a protumorigenic microenvironment.

scholarly journals 1992 Homer Smith Award. Fluid secretion, cellular proliferation, and the pathogenesis of renal epithelial cysts.

1993 ◽  
Vol 3 (12) ◽  
pp. 1841-1857 ◽  
Author(s):  
J J Grantham
Keyword(s):  
Growth Factors ◽  
Epithelial Cells ◽  
Cellular Proliferation ◽  
Early Stage ◽  
Renal Cysts ◽  
Fluid Secretion ◽  
Cyst Formation ◽  
Renal Tubules ◽  
Epidermal Growth

Renal cysts, caused by hereditary or acquired disorders, develop in tubule segments. The central pathogenetic elements of cyst formation include abnormal cellular proliferation, accumulation of intratubular liquid, and remodeling of the extracellular matrix. This review addresses the pathogenetic basis of liquid collection and cellular proliferation. Cavity liquid. At an early stage of growth, most renal cysts become detached from the tubule segment of origin; thus, transepithelial fluid secretion is the source of the liquid in most macroscopic cysts. Evidence from in situ and in vitro studies of intact cysts and epithelium cultured from cyst walls and normal renal tubules indicates that: (1) solutes (NaCl) are secreted into the cysts and water flows secondarily by osmosis; (2) active Na+ transport has a primary or secondary role in the secretion of Na+ and Cl-; and (3) the rate of liquid secretion can be modulated by hormones (arginine vasopressin), autocoids (prostaglandin E1 and E2), growth factors (epidermal growth factor), and unknown factors in cyst fluids. Cellular proliferation. Epithelial cells of renal cysts appear to proliferate more than normal. Each cyst resembles a tumor, except that the mass is composed primarily of liquid rather than cells. The proliferation of cyst epithelial cells is associated with: (1) abnormal expression of proto-oncogenes; (2) abnormal displays of morphologic and biochemical phenotypic markers; and (3) abnormal responsiveness to growth factors. The maturation arrest hypothesis, introduced as a framework to explore the pathogenetic basis of all renal cysts, supposes that the epithelial cells comprising cysts are "locked" in an immature, dedifferentiated state. Therapeutic strategies to control the growth of renal cysts may reasonably target processes that inhibit fluid secretion, maximize fluid absorption, and redifferentiate the immature and abnormally proliferative epithelial cells within cysts.

scholarly journals Immunohistological localisation of growth factors in stroma and interstitial gland tissue of goat (Capra hircus) ovary

2021 ◽  
Vol 24 (4) ◽  
pp. 478-486
Author(s):  
S. Saini ◽  
R. A. Bhat
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Stromal Cells ◽  
Transforming Growth Factor ◽  
Cell Function ◽  
Interstitial Cells ◽  
Capra Hircus ◽  
Cell Functions

The growth factors platelet derived growth factor (PDGF), transforming growth factor alpha (TGF-α) and transforming growth factor beta (TGF-β) have been demonstrated to stimulate the in vitro prolife­ration of theca and granulosa cells in different animals. The present study was conducted to localise the growth factors PDGF, TGF-α and TGF-β in different types of interstitial cells and stromal cells of normal cycling goat ovaries. Tissue fixed in formalin was processed through a graded series of alcohols and embedded in paraffin wax. The sections were immunohistochemically stained with antibo­dies against PDGF, TGF-α and TGF-β. The binding affinity of interstitial cells and stromal cells were observed and photographed. The staining pattern of PDGF, TGF-α and TGF-β was mild to strong in stromal cells. The primary and secondary interstitial cells exhibited varied staining patterns for all studied growth factors. These findings in goat suggests that PDGF, TGF-α, TGF-β were potentially an important autocrine regulator of different cell functions and possibly a paracrine regulator of ovarian cell function at various development stages.

351 EFFECTS OF FETAL CALF SERUM, GROWTH FACTOR, AND HORMONE SUPPLEMENTATION DURING IN VITRO MATURATION ON PARTHENOGENETIC ACTIVATION AND EMBRYO DEVELOPMENT OF FOLLICULAR RABBIT OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 290
Author(s):  
Z. Polgar ◽  
T. Somfai ◽  
V. Angeli ◽  
X. H. Tang ◽  
W. Ji ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Calf Serum ◽  
Complete Medium ◽  
Treatment Groups ◽  
Significant Difference

Improvement of the in vitro maturation (IVM) system for rabbit oocytes could play a role in rabbit biotechnology. Our goal was to improve IVM to have an efficient source of rabbit oocytes for further studies on nuclear transfer. The effects of FCS, growth factors, and hormone supplementation on oocyte maturation, activation, and embryo development rates were evaluated. Rabbit ovaries were transferred from the slaughterhouse to the laboratory in PBS. Oocytes were collected by aspiration and subjected to IVM in 3 types of media based on TCM-199. In one group, growth factors (50 ng mL−1 of insulin-like growth factor-I and 10 ng mL−1 of epidermal growth factor) and hormones (5 IU mL−1 of hCG and 5 IU mL−1 of pregnant mare serum gonadotropin) and BSA were added to the IVM medium (IVM+); in the other groups, IVM medium was supplemented with either 10% (IVM + 10% FCS) or 20% FCS (IVM + 20% FCS). Maturation was assessed by the presence of a polar body after 16 h. Matured oocytes were activated twice by electric stimuli (3 DC pulses, 1.6 kV cm−1, 60 µs) and twice by chemical activation (incubation with 2.5 mM 6-DMAP for 30 min, second time for 2 h) and cultured in vitro in Earle's balanced salt solution complete medium at 38.5°C under 5% CO2 in air. Cleavage rates were recorded 16 h after activation and the blastocyst rates were recorded at Day 5 of in vitro culture. Data were analyzed by ANOVA. Maturation rates did not differ between the treatment groups (Table 1). There was no significant difference in cleavage rates between the IVM+ and the IVM + 10% FCS groups; however, the cleavage rate of the IVM + 20% FCS group was significantly lower compared with the others (Table 1; P < 0.05). Development to the blastocyst did not differ significantly between the treatment groups (Table 1; P < 0.05). The results showed that high (20%) FCS supplementation during IVM had a detrimental effect on oocyte cleavage. Hormonal and growth factor supplementation had no beneficial effects on maturation, activation rates, or blastocyst formation, and in fact were not essential for in vitro embryo production in the rabbit. Table 1.Effect of hormonal and growth factor supplementation on embryo development in rabbit oocytes The project was supported by RABIOTECH OMFB-00330/2004, EU FP6 (MEXT-CT-2003-509582, and 518240), Wellcome Trust (Grant No. 070246), and TET CH-28/04.

Megakaryocytes Cultured from Patients with IMF Produced Significantly Higher mRNA but Not Protein Levels of Fibrosing Grown Factors.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3607-3607
Author(s):  
Jen-Chin Wang ◽  
Tsong H Chang ◽  
Amit Goldberg ◽  
Allan D. Novetsky ◽  
Steven Lichter ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
Growth Factor ◽  
Mrna Levels ◽  
Serum Free Medium ◽  
Free Medium ◽  
Myeloid Metaplasia ◽  
Serum Free ◽  
Protein Levels

Abstract Currently, the prevailing concept concerning the etiology of bone marrow fibrosis in patients with idiopathic myelofibrosis (IMF) is that it results from excessive production of fibrosing growth factors including transforming growth factor beta (TGF-B1), platelet-derived growth factor (PDGF), and fibroblast growth factor (FGF) from megakaryocytes and monocytes. Since megakaryocytes are difficult to isolate from bone marrow in IMF patients, this concept remains speculative. We obtained megakaryocytes (CD41+ cells) from 10-day in vitro culture of blood CD34+ cells in serum-free medium with thrombopoietin and stem cell factors as described and cultured monocytes from isolating blood CD14+ cells. Then quantitative analyses of fibrosing growth factors at the mRNA and protein levels were obtained. mRNA levels were obtained from real-time RT-PCR technique, and protein levels were obtained from ELISA analysis of the supernatant of CD41+ cells cultured 4 h in serum-free medium. The results showed 1) mRNA levels of TGF-B1, PDGF, and FGF produced by the megakaryocytes were significantly elevated in agnogenic myeloid metaplasia (AMM) compared with those in normal controls (p<0.05). While these growth factors were elevated several-fold in AMM compared with other myeloproliferative disorders (MPD) including essential thrombocythemia and polycythemia vera, they were not statistically significant. 2) mRNA levels of TGF-B1 were higher than levels of PDGF or FGF. 3) The mRNA levels of these growth factors produced from CD14+ cells were not significantly elevated in AMM compared with other MPDs or controls; the AMM mRNA levels were significantly elevated only in some patients. 4) The correlation of mRNA levels of these growth factors with the degree of myelofibrosis in AMM was significant with megakaryocytes (r=0.73) but not with monocytes (r=0.23). 5) ELISA analysis of the growth factors from the cultured megakaryocytes showed that, in most of the patients with AMM and other MPDs and in volunteer controls, the growth factors were undetectable, and only a few patients with AMM had significantly elevated protein levels of these growth factors. We conclude thatin IMF, megakaryocytes but not monocytes are the predominant cells producing fibrosing growth factors, andthe failure of finding increased protein levels of these growth factors in the in vitro system suggest that other factors are necessary to initiate translation of these growth factors in the megakaryoctes, and neutrophil emperipolesis with releasing factors may be important in this process.

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