Cultivation of post-implantation mouse and rat embryos on plasma clots

Development ◽  
1964 ◽  
Vol 12 (1) ◽  
pp. 101-111
Author(s):  
D. A. T. New ◽  
K. F. Stein
Keyword(s):  
Guinea Pig ◽  
Neural Tube ◽  
Blood Circulation ◽  
Primitive Streak ◽  
Rat Plasma ◽  
Beating Heart ◽  
Similar Degree ◽  
Rat Embryos ◽  
Mammalian Embryos ◽  
Post Implantation

Despite recent successes with the cultivation of mouse and rabbit eggs (references in Austin, 1961, pp. 144–7) techniques for the cultivation of post-implantation mammalian embryos have not hitherto advanced beyond those devised in the 1930's. Jolly & Lieure (1938) obtained development of rat and guinea-pig embryos explanted into homologous serum at stages between primitive streak and a few somites. They report that of their explanted rat embryos 37 per cent, developed an embryonic axis with a rhythmically beating heart, but only 9 per cent, a functioning circulation. None formed limb buds or a functioning allantoic circulation. Nicholas & Rudnick (1934, 1938) appear to have had a similar degree of success with rat embryos explanted into heparinized rat plasma and embryo extract. Waddington & Waterman (1933) explanted rabbit blastodiscs of primitive streak to 3-somite stages on to plasma clots; in the most successful cultures a 6–9 somite embryo was obtained with neural tube and beating heart, but without any blood circulation.

Calcium and neurulation in mammalian embryos

Development ◽  
1985 ◽  
Vol 89 (1) ◽  
pp. 1-14
Author(s):  
Martin J. Smedley ◽  
Martin Stanisstreet
Keyword(s):  
Neural Tube ◽  
Divalent Cations ◽  
Neural Cells ◽  
Cellular Mechanisms ◽  
Rat Embryos ◽  
Transmission Electron ◽  
Mammalian Embryos ◽  
Neural Ectoderm ◽  
Calcium And Magnesium

The role of calcium in neurulation in rat embryos has been studied. Rat embryos at 10·4 days of gestation, when the cephalic neural folds have elevated but not fused, have been cultured in various media, and the effects of these media on the morphology of the cephalic neural folds have been observed by scanning and transmission electron microscopy. Embryos cultured in serum containing EDTA or EGTA, or in saline without divalent cations exhibit opening, then folding back (‘collapse’) of the cephalic neural folds. The neural cells lose their elongated shape and become rounded. Older embryos in which the cephalic neural folds have already fused do not show collapse of the neural tube. Culture of 10·4-day rat embryos with elevated but unfused cephalic neural folds in calcium- and magnesium-free saline to which either calcium or magnesium has been restored shows that calcium is the divalent cation which is essential for the maintenance of the elevated neural folds. In the presence of calcium, lanthanum, which competes for calcium sites, causes opening but not collapse of the elevated cephalic neural folds. Embryos treated with trypsin show dissociation of the lateral (non-neural) ectoderm but the neural folds remain elevated. If embryos in which the cephalic neural folds have been caused to collapse are further cultured in serum the folds re-elevate, although normal neural tube morphology is not completely regained. The possible implications of these observations to the understanding of the cellular mechanisms of normal neurulation, and of neural tube malformations are discussed.

Growth of opossum embryos in vitro during organogenesis

Development ◽  
1977 ◽  
Vol 41 (1) ◽  
pp. 111-123
Author(s):  
D. A. T. New ◽  
M. Mizell ◽  
D. L. Cockroft
Keyword(s):  
Neural Tube ◽  
Blood Circulation ◽  
Primitive Streak ◽  
Yolk Sac ◽  
Body Movements

Opossum embryos, explanted between primitive streak and late fetal stages, were grown in culture for periods of 20–30 h. Many of the explants had a good heartbeat and blood circulation in embryo and yolk sac after 12 h, and a few after 24 h. Growth of the embryos included formation of the neural tube and body flexures, increase in the number of somites, differentiation of the limbs and digits, and development of the amnion and allantois. Embryos explanted during the last day of gestation showed persistent and vigorous body movements in culture, particularly of the forelimbs, head and tongue.

Whole Rat Embryos Require Methionine for Neural Tube Closure when Cultured on Cow Serum

1989 ◽  
Vol 119 (11) ◽  
pp. 1716-1725 ◽  
Author(s):  
Caroline N. D. Coelho ◽  
James A. Weber ◽  
Norman W. Klein ◽  
Willard G. Daniels ◽  
Thomas A Hoagland
Keyword(s):  
Neural Tube ◽  
Neural Tube Closure ◽  
Rat Embryos ◽  
Tube Closure

Investigation on the origin of the definitive endoderm in the rat embryo

Development ◽  
1974 ◽  
Vol 32 (2) ◽  
pp. 445-459
Author(s):  
B. Levak-Švajger ◽  
A. Švajger
Keyword(s):  
Primitive Streak ◽  
Definitive Endoderm ◽  
Rat Embryo ◽  
Chick Blastoderm ◽  
Germ Layers ◽  
Kidney Capsule ◽  
Rat Embryos ◽  
Endodermal Cells ◽  
Derivatives Of

Single germ layers (or combinations of two of them) were isolated from the primitive streak and the head-fold stage rat embryos and grown for 15 days under the kidney capsule of syngeneic adult animals. The resulting teratomas were examined histologically for the presence of mature tissues, with special emphasis on derivatives of the primitive gut. Ectoderm isolated together with the initial mesodermal wings at the primitive streak stage gave rise to tissue derivatives of all three definitive germ layers. Derivatives of the primitive gut were regularly present in these grafts. At the head-fold stage, isolated ectoderm (including the region of the primitive streak) differentiated into ectodermal and mesodermal derivatives only. Endoderm isolated at the primitive streak stage did not develop when grafted and was always completely resorbed. At the head-fold stage, however, definitive endoderm differentiated into derivatives of the primitive gut if grafted together with adjacent mesoderm. These findings indirectly suggest the migration of prospective endodermal cells from the primitive ectoderm, and therefore a general analogy with the course of events during gastrulation in the chick blastoderm.

scholarly journals Protocol for controlled modeling of human epiblast and amnion development using stem cells

2019 ◽  
Author(s):  
Yi Zheng ◽  
Jianping Fu
Keyword(s):  
Stem Cells ◽  
Primordial Germ Cells ◽  
Primitive Streak ◽  
Human Embryos ◽  
Vitro Fertilization ◽  
Human Embryology ◽  
Post Implantation ◽  
Advance Knowledge ◽  
Early Human

Abstract Due to the inaccessibility of post-implantation human embryos and the restriction on in-vitro fertilization (IVF) embryos cultured beyond 14 days, the knowledge of early post-implantation human embryogenesis remains extremely limited. Recently, we have developed a microfluidic in-vitro platform, based on human pluripotent stem cells (hPSCs), which is capable of recapitulating several key developmental landmarks of early human post-implantation embryonic development, including lumenogenesis of the epiblast (EPI), amniogenesis, and specification of primordial germ cells (PGCs) and of primitive streak (PS) cells. Given its controllability and reproducibility, the microfluidic platform provides a powerful experimental platform to advance knowledge of human embryology and reproduction. This protocol describes the preparation of the microfluidic device and its implementation for modeling human post-implantation epiblast and amnion development using hPSCs.

The T gene is necessary for normal mesodermal morphogenetic cell movements during gastrulation

Development ◽  
1995 ◽  
Vol 121 (3) ◽  
pp. 877-886 ◽  
Author(s):  
V. Wilson ◽  
L. Manson ◽  
W.C. Skarnes ◽  
R.S. Beddington
Keyword(s):  
T Cells ◽  
Neural Tube ◽  
Es Cells ◽  
Embryonic Stem ◽  
Primitive Streak ◽  
Distal Portion ◽  
Adhesion Properties ◽  
Cell Clones ◽  
Posterior Neuropore ◽  
Pass Through

The T (Brachyury) deletion in mouse is responsible for defective primitive streak and notochord morphogenesis, leading to a failure of the axis to elongate properly posterior to the forelimb bud. T/T embryonic stem (ES) cells colonise wild-type embryos, but in chimeras at 10.5 days post coitum (dpc) onwards they are found predominantly in the distal tail, while trunk paraxial and lateral mesoderm are deficient in T/T cells (Wilson, V., Rashbass, P. and Beddington, R. S. P. (1992) Development 117, 1321–1331). To determine the origin of this abnormal tissue distribution, we have isolated T/T and control T/+ ES cell clones which express lacZ constitutively using a gene trap strategy. Visualisation of T/T cell distribution in chimeric embryos throughout gastrulation up to 10.5 dpc shows that a progressive buildup of T/T cells in the primitive streak during gastrulation leads to their incorporation into the tailbud. These observations make it likely that one role of the T gene product is to act during gastrulation to alter cell surface (probably adhesion) properties as cells pass through the primitive streak. As the chimeric tail elongates at 10.5 dpc, abnormal morphology in the most distal portion becomes apparent. Comparison of T expression in the developing tailbud with the sites of accumulation of T/T cells in chimeras shows that T/T cells collect in sites where T would normally be expressed. T expression becomes internalised in the tailbud following posterior neuropore closure while, in abnormal chimeric tails, T/T cells remain on the surface of the distal tail. We conclude that prevention of posterior neuropore closure by the wedge of T/T cells remaining in the primitive streak after gastrulation is one source of the abnormal tail phenotypes observed. Accumulation of T/T cells in the node and anterior streak during gastrulation results in the preferential incorporation of T/T cells into the ventral portion of the neural tube and axial mesoderm. The latter forms compact blocks which are often fused with the ventral neural tube, reminiscent of the notochordal defects seen in intact mutants. Such fusions may be attributed to cell-autonomous changes in cell adhesion, possibly related to those observed at earlier stages in the primitive streak.

Autonomous development of parts isolated from primitive-streak-stage mouse embryos. Is development clonal?

Development ◽  
1981 ◽  
Vol 65 (Supplement) ◽  
pp. 269-287
Author(s):  
Michael H. L. Snow
Keyword(s):  
Growth Rate ◽  
Cell Proliferation ◽  
Cell Death ◽  
Primitive Streak ◽  
Mouse Embryos ◽  
Chemically Induced ◽  
Mammalian Embryos ◽  
Is Development ◽  
The Relationship ◽  
Autonomous Development

The relationship between growth rate and regionalization of amphibian, bird and mammalian embryos is briefly reviewed. In contrast to the others, mammals start gastrulation with few cells but accelerate cell proliferation coincidentally. Experiments are described which demonstrate (1) autonomous development of pieces isolated surgically from such mouse embryos, and (2) an absence of regeneration or regulation. Since such embryos regulate completely after chemically induced random cell death it is postulated that these results reflect developmental determination and a resulting mosaicism that suggests development may have a clonal basis. Maps are drawn, allocating positions to various tissues in the embryo.

scholarly journals Proteomic analysis of valproic acid-induced embryotoxicity in cultured post-implantation rat embryos

2017 ◽  
Vol 4 (1) ◽  
pp. 31-35
Author(s):  
Makoto Usami ◽  
Mina Takamatsu ◽  
Shugo Kazama ◽  
Katsuyoshi Mitsunaga ◽  
Atsuko Miyajima ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Proteomic Analysis ◽  
Rat Embryos ◽  
Post Implantation

scholarly journals Mammalian embryos show metabolic plasticity toward the surrounding environment during neural tube closure

2018 ◽  
Vol 23 (9) ◽  
pp. 794-802 ◽  
Author(s):  
Hidenobu Miyazawa ◽  
Masamichi Yamamoto ◽  
Yoshifumi Yamaguchi ◽  
Masayuki Miura
Keyword(s):  
Neural Tube ◽  
Neural Tube Closure ◽  
Metabolic Plasticity ◽  
Surrounding Environment ◽  
Mammalian Embryos ◽  
Tube Closure

THE DESTRUCTION OF O,O-DIETHYL-S-2-DIETHYLAMINOETHYL PHOSPHOROTHIOLATE BY LIVER MICROSOMES

1959 ◽  
Vol 37 (1) ◽  
pp. 297-305
Author(s):  
J. F. Scaife ◽  
D. H. Campbell
Keyword(s):  
Guinea Pig ◽  
Enzymatic Activity ◽  
Inorganic Phosphate ◽  
Enzyme System ◽  
Liver Microsomes ◽  
Rat Plasma ◽  
Fresh Tissue ◽  
Sh Groups ◽  
Diisopropyl Phosphorofluoridate ◽  
Liver Homogenates

Liver homogenates prepared from the rat, rabbit, mouse, and guinea pig possess an enzyme system capable of destroying O,O-diethyl-S-2-diethylaminoethyl phosphorothiolate at the rate of 150 to 200 μg/hr/g of fresh tissue. The homogenates prepared from the pig, dog, cow, and frog destroyed this compound at a rate of 50 to 100 μg/hr/g, but those prepared from man and the cat possessed negligible activity. Rat plasma, brain, kidney, diaphragm, whole gut, and spleen also possessed little or no activity. This enzyme system is located in the microsomes, disruption of which is accompanied by loss of enzymatic activity. The activity is dependent upon oxygen, inorganic phosphate, and diphosphopyridine nucleotide. Inhibitor studies indicate that enzymic SH-groups are necessary. The enzyme has no action on diisopropyl phosphorofluoridate, isosystox, or tetraethyl pyrophosphate, although these compounds are rapidly destroyed by the liver.

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