Response of preimplantation rat blastocysts in vitro to extracellular uterine luminal components, serum and hormones

1977 ◽  
Vol 25 (1) ◽  
pp. 265-277
Author(s):  
M.A. Surani
Keyword(s):  
Calf Serum ◽  
Giant Cells ◽  
Biologically Active ◽  
Foetal Calf Serum ◽  
Active Components ◽  
Rat Serum ◽  
Pregnant Females ◽  
Trophoblast Giant Cells

The influence of extracellular environmental factors on preimplantation rat blastocysts was tested by determining the number of embryos which escaped from their zonae pellucidae, followed by attachment and outgrowth of trophoblast giant cells, after 72 h in culture Uterine luminal ocmponents from individual females, or hormones, were included in Dulbecco's medium which contained 4 mg/ml bovine serum albumin. In about 20% of cases, uterine fluids were embryotonic. However, uterine fluids from day-5 pregnant females, the day of implantation in the rat, were more potent in these tests than uterine fluids obtained from ovariectomized females treated with progesterone alone. The potency of a mixture of the 2 fluids was also high. Uterine fluids obtained at 14 h after an injection of oestradiol and progesterone to the ovariectomized females, were also effective in these tests. Rat serum and foetal calf serum were effective too, but steroids or insulin alone in the medium had no detectable influence on embryos. Serum or uterine luminal proteins appear to be essential for maintaining the viability of the blastocysts and for inducing the responses observed here. In the uterine fluids, some proteins released into the lumen after treatment of females with oestradiol and progesterone appear to be the biologically active components. Differences in the responses of blastocysts in vitro are compared with those in vivo.

The mechanism of mouse egg-cylinder morphogenesis in vitro

Development ◽  
1981 ◽  
Vol 61 (1) ◽  
pp. 277-287
Author(s):  
A. J. Copp
Keyword(s):  
Giant Cell ◽  
Cell Formation ◽  
Cell Movement ◽  
Giant Cells ◽  
Cell Number ◽  
Dependent Change ◽  
Morphogenesis In Vitro ◽  
Trophoblast Giant Cells

The number of trophoblast giant cells in outgrowths of mouse blastocysts was determined before, during and after egg-cylinder formation in vitro. Giant-cell numbers rose initially but reached a plateau 12 h before the egg cylinder appeared. A secondary increase began 24 h after egg-cylinder formation. Blastocysts whose mural trophectoderm cells were removed before or shortly after attachment in vitro formed egg cylinders at the same time as intact blastocysts but their trophoblast outgrowths contained fewer giant cells at this time. The results support the idea that egg-cylinder formation in vitro is accompanied by a redirection of the polar to mural trophectoderm cell movement which characterizes blastocysts before implantation. The resumption of giant-cell number increase in trophoblast outgrowths after egg-cylinder formation may correspond to secondary giant-cell formation in vivo. It is suggested that a time-dependent change in the strength of trophoblast cell adhesion to the substratum occurs after blastocyst attachment in vitro which restricts the further entry of polar cells into the outgrowth and therefore results in egg-cylinder formation.

In vivo and in vitro development of mouse embryos homozygous for the embryonic lethal velvet coat (Ve) mutation

Development ◽  
1981 ◽  
Vol 66 (1) ◽  
pp. 43-55
Author(s):  
J. Rossant ◽  
K. M. Vijh
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Giant Cells ◽  
Blastocyst Stage ◽  
Parietal Endoderm ◽  
Trophoblast Giant Cells ◽  
Vitro Development ◽  
Ectoplacental Cone

Embryos homozygous for the velvet coat mutation, Ve/Ve, were recognized at 6·5 days post coitum by the reduced size of the ectodermal portions of the egg cylinder and the loose, columnar nature of the overlying endoderm. Later in development ectoderm tissues were sometimes entirely absent. Abnormalities appeared in the ectoplacental cone at 8·5 days but trophoblast giant cells and parietal endoderm appeared unaffected. Homozygotes could not be unequivocally identified at 5·5 days nor at the blastocyst stage but were recognized in blastocyst outgrowths by poor development of the inner cell mass derivatives, It has previously been suggested that Ve may exert its action at the blastocyst stage by reducing the size of the inner cell mass, but no evidence for such a reduction was found. Most of the observations on Ve/Ve homozygotes are, however, consistent with the hypothesis that Ve exerts its action primarily on later primitive ectoderm development.

Muller-cell-derived leukaemia inhibitory factor arrests rod photoreceptor differentiation at a postmitotic pre-rod stage of development

Development ◽  
1997 ◽  
Vol 124 (12) ◽  
pp. 2345-2354 ◽  
Author(s):  
C. Neophytou ◽  
A.B. Vernallis ◽  
A. Smith ◽  
M.C. Raff
Keyword(s):  
Cell Cycle ◽  
Calf Serum ◽  
Inhibitory Factor ◽  
Leukaemia Inhibitory Factor ◽  
Mouse Retina ◽  
Foetal Calf Serum ◽  
Rod Photoreceptor ◽  
Stage Of Development

In the present study, we examine rod photoreceptor development in dissociated-cell cultures of neonatal mouse retina. We show that, although very few rhodopsin+ rods develop in the presence of 10% foetal calf serum (FCS), large numbers develop in the absence of serum, but only if the cell density in the cultures is high. The rods all develop from nondividing rhodopsin- cells, and new rods continue to develop from rhodopsin- cells for at least 6–8 days, indicating that there can be a long delay between when a precursor cell withdraws from the cell cycle and when it becomes a rhodopsin+ rod. We show that FCS arrests rod development in these cultures at a postmitotic, rhodopsin-, pre-rod stage. We present evidence that FCS acts indirectly by stimulating the proliferation of Muller cells, which arrest rod differentiation by releasing leukaemia inhibitory factor (LIF). These findings identify an inhibitory cell-cell interaction, which may help to explain the long delay that can occur both in vitro and in vivo between cell-cycle withdrawal and rhodopsin expression during rod development.

Adipocytic cells cultured from marrow have osteogenic potential

1991 ◽  
Vol 99 (1) ◽  
pp. 131-139
Author(s):  
J.H. Bennett ◽  
C.J. Joyner ◽  
J.T. Triffitt ◽  
M.E. Owen
Keyword(s):  
Calf Serum ◽  
Diffusion Chamber ◽  
Osteogenic Potential ◽  
Foetal Calf Serum ◽  
Rabbit Plasma ◽  
Serum Supplement ◽  
Cells Cultured ◽  
Marrow Cells

Stromal colonies with fibroblastic morphology grown from rabbit marrow cells in culture supplemented with foetal calf serum. In this study the same marrow cells cultured with autologous rabbit plasma and hydrocortisone form colonies of a single lineage that express the adipocytic phenotype. A comparison of the potential for differentiation of cloned cell populations grown from fibroblastic and adipocytic colonies has been made using an in vivo diffusion chamber assay. The adipocytic colonies differentiated and grew to a limited size in medium with rabbit plasma and hydrocortisone, but attempts to isolate them and expand them in this medium failed. When the serum supplement was changed to foetal calf serum at day 10 the cells in the adipocytic colonies acquired a less differentiated morphology, there was a large increase in colony growth and cells were produced in sufficient numbers for the diffusion chamber assay. Thirty one fibroblastic colonies and twenty one adipocytic colonies were isolated either by limiting dilution or ring cloning and then expanded. Of these, eleven fibroblastic and eight adipocytic colonies provided enough cells (2 × 10(5) to 2 × 10(6] for implantation and culture in the chambers. Four of the eleven fibroblastic and three of the eight adipocytic colonies formed an osteogenic tissue in the chambers. It was concluded that cells that have differentiated in an adipocytic direction are able to revert to a more proliferative stage and subsequently to differentiate along the osteogenic pathway. Adipocytic and fibroblastic cells cultured in vitro from marrow have, with osteogenic cells, a common precursor in adult marrow.

Culture of mouse embryos during neurulation

Development ◽  
1981 ◽  
Vol 66 (1) ◽  
pp. 109-116
Author(s):  
T. W. Sadler ◽  
D. A. T. New
Keyword(s):  
Growth And Development ◽  
Calf Serum ◽  
Mouse Embryos ◽  
Rat Serum ◽  
Developmental Abnormalities ◽  
Time Period ◽  
Lower Protein ◽  
Made In

A comparison between static versus rotator culture systems and a variety of media (rat serum, new born calf serum, DMEM and Waymouth's) was made in an attempt to promote in vitro growth of mouse embryos from the beginning of neurulation (headfold stage) to the closure of the neural tube and formation of the limb buds (48 h). The results demonstrate that good development can be achieved for 48 h using a rotator system and that 80% of embryos cultured on rotators show growth and differentiation similar to that obtained for the same time period in vivo. Static cultures are less successful and embryos grown in this system show lower protein content and somite numbers than those maintained on rotators. Undiluted rat serum is superior to all other media tested and supports better growth and development as monitored by total protein and developmental abnormalities.

Thrombin induces Contraction And increased 51-Cr Release from Cultured Human Endothelial Cells

1979 ◽  
Author(s):  
K.S. Galdal ◽  
S.A. Evensen
Keyword(s):  
Endothelial Cells ◽  
Calf Serum ◽  
Linear Increase ◽  
Foetal Calf Serum ◽  
Human Endothelial Cells ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Test ◽  
Release Assay

Injury to human endothelial cells(EC) in primary culture was evaluated by a 51-Cr release assay, phase contrast microscopy and the trypan blue exclusion test. Normal integrity of EC, was maintained and 51-Cr release showed a slow linear increase during 24h incubation with either RPMI 1640 supplemented with 20% foetal calf serum, glutamine and antibiotics(SCM) or normal human serum(NHS). Thrombin in a concentration as low as 0.1 IU/ml induced obvious contraction of EC incubated in SCM, but the cells remained fixed to the bottom of the wells. Cell contraction was maximal after 15min and disappeared within 4h. 51-Cr release increased 2-3 fold within a few minutes and remained increased during an incubation period of 4h. The injurious effect of thrombin was inhibited in SCM containing hirudin(l.7 u/ml) or in NHS. ADP(10-5 M), endotoxin (0.1mg/ml) and 5 vasoactive agents adrenalin 5.5 10-5, noradrenalin 5.9 10-5 M, histamine 9.10-4 M, bradykinin 7.5 10-7 M and serotonin 10-5 M) did not cause cellular injury. Cultured human EC are injured by thrombin. The other tested agents do not appear to induce direct injury in vitro, but may interact with cellular elements in the blood to produce the endothelial injury previously observed in vivo.

scholarly journals Scientific basis of biomarkers and benefits of functional foods for reduction of disease risk: cancer

2002 ◽  
Vol 88 (S2) ◽  
pp. S219-S224 ◽  
Author(s):  
Joseph J. Rafter
Keyword(s):  
Cancer Risk ◽  
Functional Foods ◽  
Dietary Intervention ◽  
Disease Risk ◽  
Indirect Evidence ◽  
Biologically Active ◽  
Active Components ◽  
Vast Number

One of the most promising areas for the development of functional foods lies in modification of the activity of the gastrointestinal tract by use of probiotics, prebiotics and synbiotics. While a myriad of healthful effects have been attributed to the probiotic lactic acid bacteria, perhaps the most controversial remains that of anticancer activity. However, it must be emphasised that, to date, there is no direct experimental evidence for cancer suppression in man as a result of consumption of lactic cultures in fermented or unfermented dairy products, although there is a wealth of indirect evidence, based largely on laboratory studies. Presently, there are a large number of biomarkers available for assessing colon cancer risk in dietary intervention studies, which are validated to varying degrees. These include colonic mucosal markers, faecal water markers and immunological markers. Overwhelming evidence from epidemiological, in vivo, in vitro and clinical trial data indicates that a plant-based diet can reduce the risk of chronic disease, particularly cancer. It is now clear that there are components in a plant-based diet other than traditional nutrients that can reduce cancer risk. More than a dozen classes of these biologically active plant chemicals, now known as ‘phytochemicals’, have been identified. Although the vast number of naturally occurring health-enhancing substances appear to be of plant origin, there are a number of physiologically active components in animal products (such as the probiotics referred to above) that deserve attention for their potential role in cancer prevention.

Survival of rapidly frozen hatched mouse blastocysts

Zygote ◽  
2003 ◽  
Vol 11 (4) ◽  
pp. 361-366 ◽  
Author(s):  
Csaba Pribenszky ◽  
Sándor Cseh ◽  
Zsolt Abonyi-Tóth ◽  
László Solti
Keyword(s):  
Liquid Nitrogen ◽  
Zona Pellucida ◽  
Room Temperature ◽  
Calf Serum ◽  
Significant Loss ◽  
Rapid Freezing ◽  
Foetal Calf Serum ◽  
Freezing Medium

The objective of the present study was to examine the effect of rapid freezing on the in vitro and in vivo survival of zona-pellucida-free hatched mouse blastocysts. Hatched blastocysts were rapidly frozen in a freezing medium containing either ethylene glycol (EG) or glycerol (G) in 1.5 M or 3 M concentration. Prior to freezing, embryos were equilibrated in the freezing medium for 2 min, 10 min, 20 min or 30 min at room temperature. To freeze them, embryos were held in liquid nitrogen vapour [≈1 cm above the surface of the liquid nitrogen (LN2)] for 2 minutes and then immersed into LN2. After thawing, embryos were transferred either to rehydration medium (DPBS + 10% foetal calf serum + 0.5 M sucrose) for 10 minutes or rehydrated directly in DPBS supplemented with foetal calf serum. In vitro survival of embryos frozen with EG was higher than those frozen with G. The highest survival was obtained with 3 M EG and 2 min or 10 min equilibration prior to freezing, combined with direct rehydration after thawing. Frozen blastocysts developed into normal foetuses as well as unfrozen control ones did, with averages of 30% (control), 26% (EG) and 15% (G). The results show that hatching and hatched mouse blastocysts can be cryopreserved by a simple rapid freezing protocol in EG without significant loss of viability. Our data indicate that the mechanical protection of the zona pellucida is not needed during freezing in these stages.

Patterns of lactic dehydrogenase isozymes in mouse embryos over the implantation period in vivo and in vitro

Development ◽  
1976 ◽  
Vol 36 (3) ◽  
pp. 653-662
Author(s):  
Marilyn Monk ◽  
John Ansell
Keyword(s):  
Specific Activity ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Lactic Dehydrogenase ◽  
Giant Cells ◽  
Preimplantation Embryos ◽  
Blastocyst Implantation ◽  
Trophoblast Giant Cells

Following blastocyst implantation, or outgrowth in vitro, the LDH isozyme pattern changes from that of the maternally inherited B subunit isozyme form (LDH-1) to a pattern dominated by A subunits (Auerbach & Brinster, 1967, 1968). In preimplantation embryos we have also observed additional isozyme bands, as yet unidentified. An analysis of the pattern of newly synthesized LDH isozymes and specific activity of LDH in different regions of early post implantation embryos suggests that there is a sequential activation of A and B subunits, and that activity first appears in ICM- (inner cell mass) derived tissues and then in trophoblast-derived tissues. In vitro, in the absence of ICM cells, the transition of LDHisozyme pattern does not occur in outgrowing trophoblast giant cells. This suggests a possible inductive interaction between ICM and trophoblast.

The development of trophoblast in vitro from blastocysts containing varying amounts of inner cell mass

Development ◽  
1975 ◽  
Vol 33 (1) ◽  
pp. 177-185
Author(s):  
J. D. Ansell ◽  
M. H. L. Snow
Keyword(s):  
Giant Cell ◽  
Cell Transformation ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Calf Serum ◽  
Foetal Calf Serum ◽  
Trophoblast Cells ◽  
Intact Mouse

When intact mouse blastocysts are cultured in vitro in medium supplemented with foetal calf serum, trophoblast cells proliferate and undergo giant cell transformation such as occurs in vivo. If the amount of inner cell mass in the blastocyst is decreased by culture with [3H]-thymidine then giant cell transformation occurs normally but proliferation is reduced. In the absence of inner cell mass no proliferation occurs, and giant cell transformation is more rapid than in undamaged blastocysts.

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