Mitogenic activity of chicken chorioallantoic fluid is temporally correlated to vascular growth in the chorioallantoic membrane and related to fibroblast growth factors

Development ◽  
1991 ◽  
Vol 111 (3) ◽  
pp. 683-690
Author(s):  
I. Flamme ◽  
K. Schulze-Osthoff ◽  
H.J. Jacob
Keyword(s):  
Growth Factors ◽  
Biological Activities ◽  
Chorioallantoic Membrane ◽  
Mitogenic Activity ◽  
Fibroblast Growth ◽  
Vascular Growth ◽  
Capillary Growth ◽  
Temporally Correlated ◽  
Chorioallantoic Membrane Assay

The chorioallantoic membrane (CAM) is one of the most vascularized tissues in the chicken embryo. Capillary growth proceeds until day 10 of development and thereafter abruptly regresses. As it is generally accepted that the formation of new blood vessel is regulated by growth factors, we have investigated the presence of angiogenic and mitogenic factors in the chicken chorioallantois. In the present study, we show that chorioallantoic fluid (CAF) contains angiogenic substances that are probably synthesized in the CAM or the embryonic kidney. When applied in the chorioallantoic membrane assay, CAF from 9 day chicken embryos elicits a strong angiogenic response. This angiogenic activity of CAF is associated with pronounced mitogenic effects in vitro. Comparison of different embryonic fluids reveals that mitogenic activity is particularly evident in the CAF but not detectable in embryonic serum and amnion fluid. Expression of mitogenic activity is found to be temporally correlated with vascular growth in the CAM. High activity is detected in CAF prior to day 10 and then sharply decreases, thus preceding termination of capillary growth by one day. Heparin-sepharose affinity chromatography suggests that the biological activities of CAF probably correspond to the presence of acidic and basic fibroblast growth factor (aFGF and bFGF). In Western blot analyses of CAF, an immunoreactive bFGF-like protein of about 17 × 10(3) Mr is recognized by a monospecific anti-bFGF antiserum. This protein elutes at 2.4 M NaCl from the heparin-sepharose. The mitogenic activity of the CAF can be specifically blocked by the anti-bFGF antibody indicating bFGF to be the active mitogenic principle of the CAF. These results strongly suggest that basic and probably acidic FGF play an important role in the regulation of chorioallantoic vascular growth.

scholarly journals Growth factors are released by mechanically wounded endothelial cells.

1989 ◽  
Vol 109 (2) ◽  
pp. 811-822 ◽  
Author(s):  
P L McNeil ◽  
L Muthukrishnan ◽  
E Warder ◽  
P A D'Amore
Keyword(s):  
Endothelial Cells ◽  
Plasma Membrane ◽  
Growth Factors ◽  
Growth Factor ◽  
Mechanical Forces ◽  
Fibroblast Growth ◽  
Growth Promoting ◽  
Transient Plasma

Growth factors may be required at sites of mechanical injury and normal wear and tear in vivo, suggesting that the direct action of mechanical forces on cells could lead to growth factor release. Scraping of cells from the tissue culture substratum at 37 degrees C was used to test this possibility. We show that scraping closely mimics in vitro both the transient plasma membrane wounds observed in cells subject to mechanical forces in vivo (McNeil, P. L., and S. Ito. 1989. Gastroenterology. 96:1238-1248) and the transient plasma membrane wounds shown here to occur in endothelial cells under normal culturing conditions. Scraping of endothelial cells from the culturing substratum released into the culture medium a potent growth-promoting activity for Swiss 3T3 fibroblasts. Growth-promoting activity was released rapidly (within 5 min) after scraping but was not subsequently degraded by the endothelial cells for at least 24 h thereafter. A greater quantity of growth-promoting activity was released by cells scraped 4 h after plating than by those scraped 4 or 7 d afterwards. Thus release is not due to scraping-induced disruption of extracellular matrix. Release was only partially cold inhibitable, was poorly correlated with the level of cell death induced by scraping, and did not occur when cells were killed with metabolic poisons. These results suggest that mechanical disruption of plasma membrane, either transient or permanent, is the essential event leading to release. A basic fibroblast growth factor-like molecule and not platelet-derived growth factor appears to be partially responsible for the growth-promoting activity. We conclude that one biologically relevant route of release of basic fibroblast growth factor, a molecule which lacks the signal peptide sequence for transport into the endoplasmic reticulum, could be directly through mechanically induced membrane disruptions of endothelial cells growing in vivo and in vitro.

Embryo co-culture with bovine amniotic membrane stem cells can enhance the cryo-survival of IVF-derived bovine blastocysts comparable with co-culture with bovine oviduct epithelial cells

Zygote ◽  
2020 ◽  
pp. 1-6
Author(s):  
Shayan Nejat-Dehkordi ◽  
Ebrahim Ahmadi ◽  
Abolfazl Shirazi ◽  
Hassan Nazari ◽  
Naser Shams-Esfandabadi
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Epithelial Cells ◽  
Embryonic Development ◽  
Amniotic Membrane ◽  
Biological Activities ◽  
Survival Rates ◽  
Blastocyst Stage ◽  
Oviduct Epithelial Cells

Summary Culture conditions have a profound effect on the quality of in vitro-produced embryos. Co-culturing embryos with somatic cells has some beneficial effects on embryonic development. Considering the ability of stem cells to secrete a broad range of growth factors with different biological activities, we hypothesized that bovine amniotic membrane stem cells (bAMSCs) might be superior to bovine oviduct epithelial cells (BOECs) in supporting embryonic development and enhancing their cryo-survival. Bovine abattoir-derived oocytes were matured and fertilized in vitro. The resultant presumptive zygotes were then cultured up to the blastocyst stage in the following groups: (i) co-culture with bAMSCs, (ii) co-culture with BOECs, and (iii) cell-free culture (Con). Embryos that reached the blastocyst stage were vitrified and warmed, and their post-warming re-expansion, survival and hatching rates were evaluated after 72 h culture. Results showed that the cleavage, blastocyst, and 2 h post-warming re-expansion rates of embryos did not differ between groups. However, their survival rates in BOEC and bAMSC groups were significantly higher compared with the control (72.7, 75.6 and 37.5%, respectively, P < 0.05). In conclusion, our results showed that the cryo-survivability of IVF-derived bovine embryos could be improved through co-culturing with bAMSCs. Moreover, considering the possibility to provide multiple passages from bAMSCs compared with BOECs, due to their stemness properties and their ability to produce growth factors, the use of bAMSCs is a good alternative to BOECs in embryo co-culture systems.

Fibroblast Growth Factors Regulate Stem Cell Functioning In Vitro and in Vivo.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1705-1705
Author(s):  
Joyce S.G Yeoh ◽  
Ronald van Os ◽  
Ellen Weersing ◽  
Bert Dontje ◽  
Edo Vellenga ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Growth Factors ◽  
Cultured Cells ◽  
Cell Activity ◽  
Fibroblast Growth Factors ◽  
Fibroblast Growth ◽  
Stem Cell Activity

Abstract Fibroblast Growth Factors (FGF) are a large family of signaling molecules widely involved in tissue development, maintenance and repair. Little is known about the role of FGF/FGF-receptor signaling in the regulation of adult hematopoietic stem cells (HSC). In this study, we assessed the potential of exogenously added FGF-1/2, or retrovirally overexpressed FGF-1 to preserve HSC function in vitro and in vivo. First, we demonstrate that in vitro culture of unfractionated mouse bone marrow cells, in serum-free medium, supplemented with FGF-1 or FGF-2 or FGF-1 + 2 resulted in the robust generation of long-term repopulating (LTR) HSCs. Cultures were maintained for 12 weeks and during that time in vivo competitive reconstitution assays were performed. Stem cell activity was detectable at 3, 5, and 8 weeks after initiation of culture, but lost after 12 weeks. However, whereas 3 and 5 week cultured cells provided radioprotection in non-competitive assays, animals transplanted with 8 or 12 week cultured cells succumbed due to bone marrow failure. So far, we have been unable to expand single, highly purified Lin−Sca-1+c-Kit+ using FGF-1 + 2. Consequently, we speculated that essential intermediate cell populations or signals are required for FGF-induced stem cell conservation. To test this we cultured highly purified CD45.1 Lin−Sca-1+c-Kit+ cells in a co-culture with CD45.2 unfractionated BM. Co-cultured cells were transplanted after 5 weeks in lethally irradiated recipients, and CD45.1 chimerism levels were assessed. High levels of CD45.1 chimerism confirmed that Lin−Sca-1+c-Kit+ cells require an accessory signal in addition to FGF to induced stem cell activity in vitro. We subsequently tested stem cell potential of cells cultured in FGF-1 + 2 for 5 weeks, with the addition of SCF + IL-11 + Flt3L for the last 2, 4 or 7 days. Cell numbers increased with increasing time of growth factor presence. However, only when growth factors were present for 2 days engraftment of cultured cells in a competitive repopulation assay was increased 3.5-fold. Finally, we show by immunohistochemistry that ~10% of freshly isolated Lin−Sca-1+c-Kit+ expresses high levels of FGF-1. Retroviral overexpression of FGF-1 in stem cells resulted in increased growth potential and sustained clonogenic activity in vitro. Upon transplantation of transduced stem cells, FGF-1 overexpression resulted in increased white blood cell counts 4 weeks post-transplant compared to control animals. Most notable was a marked granulocytosis in FGF-1 overexpressing recipients Our results reveal FGF as an important regulator of HSC signaling and homeostasis. Importantly, in the presence of FGF stem cells can be maintained in vitro for 2 months. These findings open novel avenues for in vitro manipulation of stem cells for future clinical therapies.

155 IN VITRO MYELINATION NOT STIMULATED BY BRAIN FIBROBLAST GROWTH FACTORS

1980 ◽  
Vol 39 (3) ◽  
pp. 388
Author(s):  
F. J. Sell ◽  
F. C. Westall ◽  
W. R. Woodward ◽  
D. Gospodarowicz
Keyword(s):  
Growth Factors ◽  
Fibroblast Growth Factors ◽  
Fibroblast Growth

scholarly journals Modulation of Mitogenic Activity of Fibroblast Growth Factors by Inorganic Polyphosphate

2003 ◽  
Vol 278 (29) ◽  
pp. 26788-26792 ◽  
Author(s):  
Toshikazu Shiba ◽  
Daisuke Nishimura ◽  
Yumi Kawazoe ◽  
Yuichiro Onodera ◽  
Kaori Tsutsumi ◽  
...  
Keyword(s):  
Growth Factors ◽  
Fibroblast Growth Factors ◽  
Mitogenic Activity ◽  
Fibroblast Growth ◽  
Inorganic Polyphosphate

scholarly journals Nymphaeol-A Isolated from Okinawan Propolis Suppresses Angiogenesis and Induces Caspase-Dependent Apoptosis via Inactivation of Survival Signals

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Ikumi Tsuchiya ◽  
Takahiro Hosoya ◽  
Motoko Ushida ◽  
Kazuhiro Kunimasa ◽  
Toshiro Ohta ◽  
...  
Keyword(s):  
Screening Program ◽  
Biological Activities ◽  
Umbilical Vein ◽  
Chorioallantoic Membrane ◽  
Tube Formation ◽  
Mitogen Activated Protein Kinase ◽  
Cam Assay ◽  
Prenylated Flavonoids

Propolis, a resinous substance that honeybees collect to protect their beehive from enemies, is reported to have various biological activities. In our screening program to search for antiangiogenic compounds from propolis, the ethanol extracts of Okinawan propolis (EEOP) showed significant antiangiogenic activities in a tube formation assay with human umbilical vein endothelial cells (HUVECs)in vitroat 3.13 μg/mL and chorioallantoic membrane (CAM) assayin vivoat 25 μg/egg. To elucidate the active compounds of EEOP and their mode of action, we isolated some prenylated flavonoids from EEOP and found that nymphaeol-A had the strongest antiangiogenic activity among them. Nymphaeol-A significantly reducedin vivoneovessel formation in the CAM assay at 25 μg/egg. At the molecular level, nymphaeol-A markedly inactivated mitogen-activated protein kinase/ERK kinase 1/2 (MEK1/2) and extracellular signal-regulated kinase 1/2 (ERK1/2), whose molecular activations signal new vessel formation in HUVECs. In addition, nymphaeol-A dose- and time-dependently induced caspase-dependent apoptosis in tube-forming HUVECs. Taken together, nymphaeol-A was shown to inhibit angiogenesis at least in part via inactivation of MEK1/2–ERK1/2 signaling and induction of caspase-dependent apoptosis. Okinawan propolis and its major component, nymphaeol-A, may be useful agents for preventing tumor-induced angiogenesis.

139 STIMULATION OF BOVINE TROPHECTODERM MIGRATION BY FIBROBLAST GROWTH FACTORS

2010 ◽  
Vol 22 (1) ◽  
pp. 228
Author(s):  
M. I. Giassetti ◽  
Q. E. Yang ◽  
A. D. Ealy
Keyword(s):  
Cell Migration ◽  
Growth Factors ◽  
Fibroblast Growth Factors ◽  
Large Family ◽  
Lower Surface ◽  
Epifluorescence Microscopy ◽  
Receptor Subtypes ◽  
Fibroblast Growth ◽  
Conceptus Development

Following hatching, bovine and ovine blastocysts elongate into tubular and then filamentous conceptuses that remain free-floating for several days before attaching to the uterine lining. Elongation is marked by trophectoderm proliferation and changes in trophectoderm shape. The ultimate goal of this work is to identify uterine- and conceptus-derived factors that control peri-attachment conceptus development in cattle. Fibroblast growth factors (FGF) encompass a large family of mitogens, morphogens, and angiogenic factors produced by various tissues, including the bovine/ovine endometrium and conceptus. FGF2 and FGF10 are of particular interest because uterine production of FGF2 and conceptus production of FGF10 intensify as elongation takes place in cattle and sheep. The objective of this work was to determine if FGF2 and FGF10 stimulate bovine trophectoderm migration during culture. Migration assays were conducted with CT1 cells, a trophectoderm line established from a bovine in vitro-produced blastocyst outgrowth. Cells were seeded on 8-μm pore Transwell inserts (Corning Inc., Corning, NY, USA; 50,000 cells/insert) and submerged in serum-free DMEM containing treatments (0, 0.5, 5, 50, and 500 ng mL-1 of recombinant bovine FGF2 or human FGF10). After 12 h, cells that migrated onto the lower surface were fixed, stained, and processed for counting using epifluorescence microscopy. Migrated cells were counted in 5 non-overlapping locations on each of 4 replicate Transwell inserts for each treatment. Experiments were repeated on at least 3 different occasions. Analysis of variance was completed. Differences in individual means were partitioned further by completing pair-wise comparisons. Supplementation with 5 or 50 ng mL-1 of FGF2 increased (P = 0.06 and P = 0.002, respectively) migration of CT1 cells when compared with controls (327 ± 17 or 485 ± 40 cells, respectively, v. 162 ± 16 cells). Supplementation with 500 ng mL-1 of FGF2 further increased (P < 0.02) migration when compared with controls and cells exposed to lower levels of FGF2 (548 ± 116 cells). FGF10 also stimulated CT1 migration. Supplementation with 0.5 ng mL-1 of FGF10 increased (P = 0.06) cell migration v. controls (254 ± 48 v. 184 ± 24 cells). Supplementation with 5 ng mL-1 further increased (P < 0.007) cell migration (373 ± 29 cells). Exposure to greater FGF10 concentrations did not further enhance cell migration. To summarize, both FGF2 and FGF10 promoted CT1 migration, suggestive of a potential function in regulating trophectoderm development, differentiation, and/or morphogenesis during peri-attachment conceptus development. FGF10 appeared to be more potent than FGF2 at mediating CT1 migration. The reason for this disparity has not been resolved but likely involves differences in ligand affinities to certain receptor subtypes. This project was supported by NRI Competitive Grant No. 2008-35203-19106 from the USDA-CSREES.

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