Preimplantation mouse embryos cultured in mouse,rat and human sera: differentiation and sister chromatid exchange

Summary Development of 4-cell and of 8-cell mouse embryos and of morulae and blastocysts is inhibited in vitro by mouse serum but not by rat or human sera which also do not influence sister chromatid exchange in cultured morulae and blastocysts.     

Effects of ethanol and acetaldehyde on cultured pre-implantation mouse embryos

Pre-implantation 2-cell stage mouse embryos, obtained from superovulated CF-1 mice, were exposed to ethanol and acetaldehyde through the culture medium for 60 min followed by a 105-h incubation period. Control and ethanol exposed embryos survived equally well in ethanol concentrations as high as 800 mg/100 ml medium and acetaldehyde levels up to 10 mg/100 ml medium.     

Effects of ethanol and acetaldehyde on cultured pre-implantation mouse embryos

Summary Pre-implantation 2-cell stage mouse embryos, obtained from superovulated CF-1 mice, were exposed to ethanol and acetaldehyde through the culture medium for 60 min followed by a 105-h incubation period. Control and ethanol exposed embryos survived equally well in ethanol concentrations as high as 800mg/100 ml medium and acetaldehyde levels up to 10 mg/100 ml medium.     

Comparison of the in vitro development of mouse single blastomeres with and without the zona pellucida

No difference was observed during in vitro development between mouse single blastomeres with and without the zona pellucida, isolated from 2- and 4-cell stage eggs.     

Comparison of the in vitro development of mouse single blastomeres with and without the zona pellucida

Summary No difference was observed during in vitro development between mouse single blastomeres with and without the zona, pellucida, isolated from 2- and 4-cell stage eggs.     

Close association of erythrocytes with surface of late mouse blastocysts: SEM analysis

SEM analysis of preimplantation mouse blastocysts reveals closely associated red blood cells on involuted surfaces of trophoblasts, and indication of the capability for initial phases of phagocytosis at an early developmental stage.     

The effect of cyclic cytidine 3',5'-monophosphate (cCMP) on the in vitro development, hatching and attachment of the mouse blastocyst

The in vitro development and attachment of hatched mouse blastocysts on the untreated substratum was enhanced by 10 microM dibutyryl cCMP (dbcCMP). The result suggests that cCMP has an effect on embryonic development and on the blastocyst attachment process.     

Close association of erythrocytes with surface of late mouse blastocysts: SEM analysis

Summary SEM analysis of preimplantation mouse blastocysts reveals closely associated red blood cells on involuted surfaces of trophoblasts, an indication of the capability for initial phases of phagocytosis at an early developmental stage.Acknowledgment. This study was supported by NICHD grant HD-06234 and NIH grant RR-05384. Author for reprint requests.     

Effects of low temperatures on survival of frozen-thawed mouse embryos

Summary Almost no damage to mouse morulae was observed between 0 and –40°C, and freezing damage to embryos in DMSO, glycerol or ethylene glycol occurred after exposure to –60, –50 or –50°C, respectively. Cooling embryos in DMSO or glycerol to temperatures below –50 to –60°C increased freezing damage. To the contrary, in the presence of ethylene, glycol, no more damage occurred after exposure to temperatures below –50°C.     

Effects of the temperature of ice-seeding on survival of frozen-and-thawed mouse morulae

Summary Mouse morulae were frozen to –196°C in the presence of 1.2 M ethylene glycol or 1 M glycerol. After iceseeding in embryo samples at –10°C or below, the survival rates in vitro were lower than those obtained by ice-seeding between –4° and –8°C. The proportion morulae developing into live young in vivo after ice-seeding at –13°C and freezing was lower than that of embryos seeded at –4°C.     

Effect of inhibitors of polyamine biosynthesis on primary differentiation of mouse egg]

Alpha-Methylornithine, an inhibitor of the synthesis of putrescine does not affect the cleavage of Mouse eggs, cultured in vitro from the 2-cell stage, before blastocyst formation, whereas methylglyoxal-Bis (guanylhydrazone), an inhibitor of the syntheses of spermidine and spermine induces the embryos to become quiescent at about the 8-cell stage; resumption of their development, after transfer to fresh medium, is followed by a delay in cavitation. These results may be related to the biological clock theory for primary differentiation.     

Progesterone induced diminished incorporation of exogenous14C-amino acids in diapausing mouse blastocysts

Summary Under identical in vitro labeling conditions, significantly lower amounts of radioactivity were found a) in proteins of early diapausing blastocysts (days 7–9 p.c.) vs normal, late blastocysts (day 5 p.c.) and b) in proteins of days 19–23 p.c. diapausing blastocysts from ovariectomized mothers treated with progesterone vs similar embryos from untreated mothers. Thus, progesterone, which maintains the viability of embryos during prolonged diapause, causes diminished utilization of exogenous amino acids for protein synthesis in these embryos.Acknowledgment. This study was supported, in part, by NICHD grant HD-06234.     

Gene-manipulated embryonic stem cells for rat transgenesis

Embryonic stem cells (ESCs) are derived from blastocysts and are capable of differentiating into whole tissues and organs. Transplantation of ESCs into recipient blastocysts leads to the generation of germline-competent chimeras in mice. Transgenic, knockin, and knockout gene manipulations are available in mouse ESCs, enabling the production of genetically modified animals. Rats have important advantages over mice as an experimental system for physiological and pharmacological investigations. However, in contrast to mouse ESCs, rat ESCs were not established until 2008 because of the difficulty of maintaining pluripotency. Although the use of signaling inhibitors has allowed the generation of rat ESCs, the production of genetically modified rats has been difficult due to problems in rat ESCs after gene introduction. In this review, we will focus on some well-documented examples of gene manipulation in rat ESCs.     

Mouse egg cylinders developed in vitro may form benign and malignant teratoid tumors

Summary Egg cylinders developed in vitro from explanted mouse blastocysts were transplanted under the kidney capsules of adult mice to assess their developmental potential. In vitro grown egg cylinders are capable of differentiation into somatic tissues and may give rise to stem cells of teratocarcinoma like the developmentally equivalent 7 day mouse embryos grown in vivo.     

Growth of trophoblast in mouse lung

Summary When blastocysts are introduced into the lungs of mice via the circulation, trophoblast cells grow, as they do in other extrauterine sites.     

Periimplantation-like growth and development of mouse blastocysts in medium containing horse serum

Horse serum (HS) supported growth and differentiation of blastocysts as well as or better than fetal bovine serum (FBS) (p less than 0.01) as measured by a) the onset of trophoblastic outgrowth, b) the size of the resultant outgrowths, c) the size of nuclei in trophoblasts after their outgrowth and d) the size of the inner cell mass in outgrowths. Moreover, polyamines were found to be approximately 10-100 times less toxic to embryos when added to medium containing HS than when the medium contained FBS.     

The effect of cyclic cytidine 3′,5′-monophosphate (cCMP) on the in vitro development,hatching and attachment of the mouse blastocyst

Summary The in vitro development and attachment of hatched mouse blastocysts on the untreated substratum was enhanced by 10 M dibutyryl cCMP (dbcCMP). The result suggests that cCMP has an effect on embryonic development and on the blastocyst attachment process.25 November1986     

The effects of time of equilibration with cryoprotectants at 0 degree C prior to freezing on the survival of mouse embryos frozen by the two-step method

Mouse embryos were frozen by the two-step method after equilibration for 0.1-60 min with cryoprotectants at 0 degree C. No survival or a very low survival was obtained after equilibration for only 0.1 min. The morulae showed the highest survival rates when equilibration time was 5-30 min with 2 M DMSO, 20-30 min with 2 M glycerol, 5-10 min with 2 M ethylene glycol and 20-30 min with 2 M propylene glycol, respectively.     

Solution structure and activity of mouse lysozyme M

The three-dimensional structure of mouse lysozyme M, glycoside hydrolase, with 130 amino acids has been determined by heteronuclear NMR spectroscopy. We found that mouse lysozyme M had four alpha-helices, two 3(10)helices, and a double- and a triple-stranded anti-parallel beta-sheet, and its structure was very similar to that of hen lysozyme in solution and in the crystalline state. The pH activity profile of p-nitrophenyl penta N-acetyl-beta-D-chitopentaoside hydrolysis by mouse lysozyme M was similar to that of hen lysozyme, but the hydrolytic activity of mouse lysozyme M was lower. From analyses of binding affinities of lysozymes to a substrate analogue and internal motions of lysozymes, we suggest that the lower activity of mouse lysozyme M was due to the larger dissociation constant of its enzyme-substrate complex and the restricted internal backbone motions in the molecule.