scholarly journals Granulocyte-Macrophage Colony Stimulating Factor in Single Blastocyst Conditioned Medium as a Biomarker for Predicting Implantation Outcome of Embryo

2021 ◽  
Vol 12 ◽  
Author(s):  
Peilin Chen ◽  
Chunyu Huang ◽  
Qing Sun ◽  
Huixian Zhong ◽  
Feng Xiong ◽  
...  
Keyword(s):  
Conditioned Medium ◽  
Pregnancy Outcome ◽  
Embryo Quality ◽  
Colony Stimulating Factor ◽  
Developmental Potential ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Selection Of

BackgroundIt is highly desirable to develop new strategies based on secretomics to more accurately selection of embryos with the highest developmental potential for transfer. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been reported to promote embryo development and pregnancy establishment. However, the predictive value of GM-CSF in single blastocyst selection remains unclear. This study is to determine the concentration of GM-CSF in human single-blastocyst conditioned medium (SBCM) and to evaluate its association with embryo quality and pregnancy outcome.MethodsThe patients with ≤38 years of age receiving the first cycle of assisted reproductive therapy were included in this study. The patients who had <4 top-quality embryos formed by the fertilized two pronuclear zygotes on day 3 were excluded. A total of 126 SBCM samples (SBCMs) were included, of which blastocysts from 77 SBCMs were later transferred in subsequent frozen-thawed embryo transfer. The concentrations of GM-CSF were detected by single-molecule array (SIMOA) and analyzed for their possible association with embryo quality and pregnancy outcomes. The top-quality embryo (TQ), positive HCG (HP), clinical pregnancy (CP), and ongoing pregnancy (OP) rates were determined and compared between groups divided based on GM-CSF concentrations.ResultsThe detection rate of GM-CSF was found to be 50% in all SBCMs. There were significant differences in TQ rate, HP rate, CP rate and OP rate among high concentration group, medium concentration group and low concentration group. Both GM-CSF alone or GM-CSF combined with the morphological score (MS) had a greater AUC of ROC curve than that of MS alone to predict the pregnancy outcome, and GM-CSF combined with MS had the highest AUC.ConclusionsThe concentration of GM-CSF in SBCM was detected at fg/ml levels, which was associated with embryo quality and pregnancy outcome. Collectively, GM-CSF may be used as a biomarker for prediction of pregnancy outcome and selection of embryos with high developmental potential for transfer in assisted reproductive technology (ART).

scholarly journals Granulocyte/macrophage colony-stimulating factor is essential for the viability and function of cultured murine epidermal Langerhans cells.

1987 ◽  
Vol 166 (5) ◽  
pp. 1484-1498 ◽  
Author(s):  
M D Witmer-Pack ◽  
W Olivier ◽  
J Valinsky ◽  
G Schuler ◽  
R M Steinman
Keyword(s):  
Dendritic Cells ◽  
Conditioned Medium ◽  
Langerhans Cells ◽  
Cell Mediated Immunity ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Accessory Function ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

A panning method has been developed to enrich Langerhans cells (LC) from murine epidermis. In standard culture media, the enriched populations progressively lose viability over a 3-d interval. When the cultures are supplemented with keratinocyte-conditioned medium, LC viability is improved and the cells increase in size and number of dendritic processes. Accessory function, as monitored by stimulating activity in the mixed lymphocyte reaction (MLR), increases at least 10-20-fold. The conditioned media of stimulated macrophages and T cells also support the viability and maturation of cultured LC. A panel of purified cytokines has been tested, and only granulocyte/macrophage colony-stimulating factor (GM-CSF) substitutes for bulk-conditioned medium. The recombinant molecule exhibits half-maximal activity at 5 pM. Without activity are: IL-1-4; IFN-alpha/beta/gamma; cachectin/TNF; M- and G-CSF. A rabbit anti-GM-CSF specifically neutralizes the capacity of keratinocyte-conditioned medium to generate active LC. However, GM-CSF is not required for LC function during the MLR itself. We conclude that the development of immunologically active LC in culture is mediated by GM-CSF. The observation that these dendritic cells do not respond to lineage-specific G- and M-CSFs suggests that LC represent a distinct myeloid differentiation pathway. Because GM-CSF can be made by nonimmune cells and can mediate the production of active dendritic cells, this cytokine provides a T-independent mechanism for enhancing the sensitization phase of cell-mediated immunity.

scholarly journals Human serum megakaryocyte colony-stimulating activity appears to be distinct from interleukin-3, granulocyte-macrophage colony-stimulating factor, and lymphocyte-conditioned medium

Blood ◽  
1990 ◽  
Vol 76 (2) ◽  
pp. 290-297 ◽  
Author(s):  
EM Mazur ◽  
JL Cohen ◽  
J Newton ◽  
P Sohl ◽  
A Narendran ◽  
...  
Keyword(s):  
Human Serum ◽  
Conditioned Medium ◽  
Colony Growth ◽  
Colony Stimulating Factor ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Colony Stimulating Activity ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Sera from patients with bone marrow megakaryocyte aplasia are a rich source of megakaryocyte colony-stimulating activity (Meg-CSA). Other biologic materials exhibiting Meg-CSA include phytohemagglutinin- stimulated human lymphocyte-conditioned medium (PHA-LCM), recombinant interleukin-3 (IL-3), and recombinant granulocyte macrophage colony- stimulating factor (GM-CSF). Neutralizing antisera to both recombinant IL-3 and GM-CSF were used to evaluate the relationship among these sources of Meg-CSA. Varying dilutions of IL-3 and GM-CSF antisera were tested in plasma clot cultures of normal human peripheral blood megakaryocyte progenitors optimally stimulated by either IL-3 (1 U/mL), GM-CSF (1 U/mL), PHA-LCM (2.5% to 5% vol/vol), or aplastic human serum (10% vol/vol). IL-3 antiserum at dilutions up to 1/2,000 totally abrogated megakaryocyte colony growth stimulated by IL-3. A 1/500 dilution of GM-CSF antiserum completely eliminated GM-CSF-induced megakaryocyte colony development. A combination of anti-IL-3 and anti- GM-CSF, each at a 1/500 dilution, inhibited all megakaryocyte colony growth stimulated by optimal concentrations of IL-3 and GM-CSF together. There was no neutralizing crossreactivity between the IL-3 and GM-CSF antisera. At maximally neutralizing concentrations, IL-3 antiserum inhibited 66% of the megakaryocyte colony growth stimulated by PHA-LCM. Residual megakaryocyte colony growth was eliminated by the addition of a 1/500 dilution of anti-GM-CSF.

scholarly journals Granulocyte/macrophage colony-stimulating factor is an intrinsic keratinocyte-derived growth factor for human melanocytes in UVA-induced melanosis

1996 ◽  
Vol 313 (2) ◽  
pp. 625-631 ◽  
Author(s):  
Genji IMOKAWA ◽  
Yukihiro YADA ◽  
Mitsutoshi KIMURA ◽  
Naoko MORISAKI
Keyword(s):  
Dna Synthesis ◽  
Conditioned Medium ◽  
Binding Sites ◽  
Human Keratinocytes ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Human Melanocytes ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Recently we demonstrated that endothelins secreted from human keratinocytes act as intrinsic mitogens and melanogens for human melanocytes in UVB-induced melanosis. We show here that UVA-induced melanosis is associated with other keratinocyte-derived growth factors, secretion of which is specifically stimulated after exposure of human keratinocytes to UVA. Medium conditioned by UVA-exposed human keratinocytes elicited a significant increase in DNA synthesis by cultured human melanocytes in a UVA dose-dependent manner. Analysis of endothelin-1 and interleukin (IL)-1α in the conditioned medium by ELISA, both of which are major keratinocyte-derived cytokines involved in UVB-associated melanocyte activation, revealed that UVA exposure did not cause human keratinocytes to stimulate the secretion of the two cytokines. In contrast, the levels of several other cytokines such as IL-6, IL-8 and granulocyte/macrophage colony-stimulating factor (GM-CSF) were significantly increased in the conditioned medium of human keratinocytes after exposure to UVA at a dose of 1.0 J/cm2. The gel chromatographic profile of UVA-exposed keratinocyte-conditioned medium demonstrated that there were two factors (P-1 and P-2) with molecular masses of approx. 20 and 1 kDa respectively that stimulate DNA synthesis in human melanocytes, and the larger species (P-1) also increased melanization as assessed by [14C]thiouracil incorporation. Quantitative analysis of cytokines in chromatographic fractions by ELISA revealed the P-1 fraction to be consistent with the molecular mass profile of GM-CSF. Furthermore the stimulatory effect of the P-1 fraction on DNA synthesis in human melanocytes was neutralized by antibodies to GM-CSF, but not to basic fibroblast growth factor or stem cell factor. Binding and proliferation assays with recombinant GM-CSF demonstrated that human melanocytes possess specific binding sites for GM-CSF(Kd 2.11 nM; binding sites, 2.5-3.5×104 per cell), and recombinant GM-CSF at concentrations of more than 10 nM significantly stimulated DNA synthesis and melanization. These findings suggest that GM-CSF secreted by keratinocytes plays an essential role in the maintenance of melanocyte proliferation and UVA-induced pigmentation in the epidermis.

scholarly journals Human serum megakaryocyte colony-stimulating activity appears to be distinct from interleukin-3, granulocyte-macrophage colony-stimulating factor, and lymphocyte-conditioned medium

Blood ◽  
1990 ◽  
Vol 76 (2) ◽  
pp. 290-297
Author(s):  
EM Mazur ◽  
JL Cohen ◽  
J Newton ◽  
P Sohl ◽  
A Narendran ◽  
...  
Keyword(s):  
Human Serum ◽  
Conditioned Medium ◽  
Colony Growth ◽  
Colony Stimulating Factor ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Colony Stimulating Activity ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Sera from patients with bone marrow megakaryocyte aplasia are a rich source of megakaryocyte colony-stimulating activity (Meg-CSA). Other biologic materials exhibiting Meg-CSA include phytohemagglutinin- stimulated human lymphocyte-conditioned medium (PHA-LCM), recombinant interleukin-3 (IL-3), and recombinant granulocyte macrophage colony- stimulating factor (GM-CSF). Neutralizing antisera to both recombinant IL-3 and GM-CSF were used to evaluate the relationship among these sources of Meg-CSA. Varying dilutions of IL-3 and GM-CSF antisera were tested in plasma clot cultures of normal human peripheral blood megakaryocyte progenitors optimally stimulated by either IL-3 (1 U/mL), GM-CSF (1 U/mL), PHA-LCM (2.5% to 5% vol/vol), or aplastic human serum (10% vol/vol). IL-3 antiserum at dilutions up to 1/2,000 totally abrogated megakaryocyte colony growth stimulated by IL-3. A 1/500 dilution of GM-CSF antiserum completely eliminated GM-CSF-induced megakaryocyte colony development. A combination of anti-IL-3 and anti- GM-CSF, each at a 1/500 dilution, inhibited all megakaryocyte colony growth stimulated by optimal concentrations of IL-3 and GM-CSF together. There was no neutralizing crossreactivity between the IL-3 and GM-CSF antisera. At maximally neutralizing concentrations, IL-3 antiserum inhibited 66% of the megakaryocyte colony growth stimulated by PHA-LCM. Residual megakaryocyte colony growth was eliminated by the addition of a 1/500 dilution of anti-GM-CSF.

scholarly journals Toxoplasma gondii Induces Granulocyte Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Secretion by Human Fibroblasts: Implications for Neutrophil Apoptosis

2002 ◽  
Vol 70 (11) ◽  
pp. 6048-6057 ◽  
Author(s):  
Jacqueline Y. Channon ◽  
Kristin A. Miselis ◽  
Laurie A. Minns ◽  
Chaitali Dutta ◽  
Lloyd H. Kasper
Keyword(s):  
Conditioned Medium ◽  
Human Fibroblasts ◽  
Neutrophil Apoptosis ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Soluble Mediator ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

ABSTRACT Human neutrophils are rescued from apoptosis following incubation with once-washed, fibroblast-derived Toxoplasma gondii tachyzoites. Both infected and uninfected neutrophils are rescued, implicating a soluble mediator. In this study we investigated the origin and identity of this soluble mediator. Neutrophils were incubated either with purified tachyzoites or with conditioned medium derived from T. gondii-infected human fibroblasts. Conditioned medium was found to be a potent stimulus that delayed neutrophil apoptosis up to 72 h, whereas purified and extensively washed tachyzoites had no effect. Delayed apoptosis correlated with up-regulation of the neutrophil antiapoptotic protein, Mcl-1, and the neutrophil interleukin 3 receptor α subunit (IL-3Rα), suggesting a role for granulocyte-macrophage colony-stimulating factor (GM-CSF). GM-CSF and granulocyte colony-stimulating factor (G-CSF) were measurable in conditioned medium by enzyme-linked immunosorbent assay. Neutralizing antibodies to GM-CSF and G-CSF were additive in abrogating delayed neutrophil apoptosis induced by conditioned medium. Inhibitors of Src family tyrosine kinases, Gi proteins, phosphatidylinositol 3-kinase, p44 erk1 and p42 erk2 mitogen-activated protein kinases, and Jak2 kinases partially attenuated the effect of conditioned medium, consistent with a role for G-CSF and/or GM-CSF. Hence, delayed neutrophil apoptosis is mediated by GM-CSF and G-CSF secreted by T. gondii-infected human fibroblasts. This enhanced neutrophil survival may contribute to the robust proinflammatory response elicited in the T. gondii-infected host.

scholarly journals T cells from eosinophilic patients produce interleukin-5 with interleukin-2 stimulation

Blood ◽  
1989 ◽  
Vol 73 (7) ◽  
pp. 1809-1813 ◽  
Author(s):  
H Enokihara ◽  
S Furusawa ◽  
H Nakakubo ◽  
H Kajitani ◽  
S Nagashima ◽  
...  
Keyword(s):  
T Cells ◽  
Conditioned Medium ◽  
Colony Formation ◽  
Interleukin 2 ◽  
Colony Stimulating Factor ◽  
Interleukin 5 ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Anti-murine (m) interleukin-5 (IL-5) antibody was found to inhibit eosinophil (Eo) colony formation stimulated by recombinant human (rh) IL-5, but did not inhibit the production of Eo stimulated by rh IL-3 or granulocyte-macrophage colony-stimulating factor (GM-CSF). Conditioned medium (CM) prepared from eosinophilic patients' T cells with interleukin-2 (IL-2) stimulation (T-IL-2-CM), was found to contain CFU- Eo growth-stimulating factor. Using anti-mIL-5 antibody, we demonstrated that T-IL-2-CM from patients with eosinophilia contained a significant amount of IL-5. We also detected IL-5 mRNA in T cells from eosinophilic patients with IL-2 stimulation. These results suggest that IL-5 plays an important role in the induction of selective eosinophilia in humans and that IL-5 is produced from T cells with IL-2 stimulation.

scholarly journals T cells from eosinophilic patients produce interleukin-5 with interleukin-2 stimulation

Blood ◽  
1989 ◽  
Vol 73 (7) ◽  
pp. 1809-1813
Author(s):  
H Enokihara ◽  
S Furusawa ◽  
H Nakakubo ◽  
H Kajitani ◽  
S Nagashima ◽  
...  
Keyword(s):  
T Cells ◽  
Conditioned Medium ◽  
Colony Formation ◽  
Interleukin 2 ◽  
Colony Stimulating Factor ◽  
Interleukin 5 ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Anti-murine (m) interleukin-5 (IL-5) antibody was found to inhibit eosinophil (Eo) colony formation stimulated by recombinant human (rh) IL-5, but did not inhibit the production of Eo stimulated by rh IL-3 or granulocyte-macrophage colony-stimulating factor (GM-CSF). Conditioned medium (CM) prepared from eosinophilic patients' T cells with interleukin-2 (IL-2) stimulation (T-IL-2-CM), was found to contain CFU- Eo growth-stimulating factor. Using anti-mIL-5 antibody, we demonstrated that T-IL-2-CM from patients with eosinophilia contained a significant amount of IL-5. We also detected IL-5 mRNA in T cells from eosinophilic patients with IL-2 stimulation. These results suggest that IL-5 plays an important role in the induction of selective eosinophilia in humans and that IL-5 is produced from T cells with IL-2 stimulation.

217 EFFECTS OF PORCINE GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR ON PORCINE IN VITRO-FERTILIZED EMBRYOS

2011 ◽  
Vol 23 (1) ◽  
pp. 207
Author(s):  
S. S. Kwak ◽  
D. Biswas ◽  
S. H. Hyun
Keyword(s):  
Polyvinyl Alcohol ◽  
Colony Stimulating Factor ◽  
Blastocyst Formation ◽  
Cleavage Rate ◽  
Developmental Potential ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

The cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) is expressed in the female reproductive tract and is one of the regulatory molecules that mediate maternal effects on the growth and development of pre-implantation embryos in several species. The objective of the present study was to investigate the effects of porcine GM-CSF (pGM-CSF) on the developmental potential of porcine IVF embryos. All experiments were performed with zygotes that were produced in vitro and cultured in porcine zygote medium-3–polyvinyl alcohol-based medium. Data were analysed with PASW statistics-17 (SPSS Inc., Chicago, IL) using Duncan’s multiple range test. A total 865 zygotes in 4 replicates were used with different concentrations of pGM-CSF (0, 2, 10, 100 ng mL–1) in Experiment 1. It was demonstrated that 10 ng mL–1 of pGM-CSF could increase (15.1 ± 2.2) blastocyst development significantly (P < 0.05) compared with the control (6.1 ± 0.7). There was no effect on cleavage rate. In blastocyst formation, early and expanded blastocysts were significantly (P < 0.05) higher in the 10 ng mL–1 of pGM-CSF group compared with the control. In Experiment 2, a total 839 zygotes with at least 5 replicates in each group were used, and whether pGM-CSF would act to increase blastocyst yield before or after Day 4 development was tested. Zygotes were cultured with the following treatments: 1) zygotes cultured with fresh porcine zygote medium–polyvinyl alcohol medium from Days 0 to 7 post-insemination as a control; 2) medium supplemented with 10 ng mL–1 of pGM-CSF from Days 0 to 4 followed by no pGM-CSF from Days 4 to 7; 3) medium alone from Days 0 to 4 followed by supplementation with 10 ng mL–1 of pGM-CSF from Days 4 to 7; and 4) medium supplemented with 10 ng mL–1 of pGM-CSF from Days 0 to 7. As compared with the controls (7.8 ± 0.7), pGM-CSF influenced the percentage of blastocyst formation when pGM-CSF was added from Days 4 to 7 (14.6 ± 1.6) or Days 0 to 7 (15.2 ± 1.8), but not from Days 0 to 4 (8.7 ± 1.5). Similarly, the early blastocyst formation rates were significantly higher in the Day 4 to 7 culture period compared with the control, and expanded blastocyst formation was significantly higher in the Day 4 to 7 and Day 0 to 7 culture periods. There was no significant different in cleavage rate among these groups. In conclusion, these data suggest that supplementation of pGM-CSF in in vitro culture medium at Days 4 to 7 or Days 0 to 7 promotes the developmental potential of porcine IVF embryos. This work was supported by a grant (20070301034040) from the BioGreen 21 Program, Rural Development Administration, Republic of Korea.

scholarly journals Granulocyte/macrophage colony-stimulating factor from mouse lung conditioned medium. Purification of multiple forms and radioiodination

1986 ◽  
Vol 235 (3) ◽  
pp. 805-814 ◽  
Author(s):  
A W Burgess ◽  
D Metcalf ◽  
L G Sparrow ◽  
R J Simpson ◽  
E C Nice
Keyword(s):  
Conditioned Medium ◽  
Cell Surface Receptor ◽  
Mouse Lung ◽  
Mouse Bone Marrow ◽  
Colony Stimulating Factor ◽  
Binding Studies ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Four forms of mouse granulocyte/macrophage colony-stimulating factor (GM-CSF) were purified 100,000-fold from mouse lung conditioned medium. Each of the CSF species stimulated the formation of both granulocyte and macrophage colonies, and half-maximal stimulation in the semi-solid mouse bone-marrow colony assay occurred at 1 pm. The four GM-CSF species exhibited similar charge microheterogeneity, focusing between pH 4.2 and pH 5.2. On SDS/polyacrylamide gels two of the GM-CSF sub-species had apparent Mr values of 23,000, and the other two, 21, 000. Treatment with neuraminidase decreased the Mr values of these two sets to 21,000 and 19,000 respectively. Incubation with endoglucosidase F decreased the charge heterogeneity and the Mr of all species to 16,500. A gas-phase radioiodination procedure was used to incorporate 2-3 atoms of 125I/molecule into purified GM-CSF without any loss of biological activity. The 125I-labelled GM-CSF was analysed on a microbore reversed-phase h.p.l.c. column to determine its specific radioactivity directly. This 125I-labelled GM-CSF molecule is suitable for cell-surface receptor-binding studies.

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