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Biomedical subjects

C Polge

Publications and source records attributed to C Polge.

At least 19 recordsLinked to original sources
PubMed1989

Survival of mouse embryos after being frozen in glycerol-sucrose mixture.

Random bred female albino mice (6-8 weeks old) were used as a source of embryos. 8- to 16 cell embryos were dehydrated in glycerol-sucrose mixture in 0.25 ml straws at room temperature. Straws were cooled at the rate of 5 degrees C/min to -7 degrees C. Seeding was induced by touching the out side of the straw at -7 degrees C. Straws were further cooled at 0.5 degree C/min down to -35 degrees C and then plunged into liquid N2. Thawing of straws was done by direct transfer into water at 35 degrees C. Frozen-thawed embryos were cultured in a CO2 incubator maintained at 39 degrees C. Out 190 embryos (8-16 cell) initially frozen, 169 (88.94%) were recovered on thawing. 158 (93.5%) out of 169 were apparently normal and used for culture. 75 (47.46%) developed to morulae/early blastocysts and 72 (45.56%) to expanded blastocysts on 24 and 48 hr culture respectively. In conclusion, the incorporation of sucrose in the freezing medium at a concentration of 0.25 M has led us to propose a freezing, thawing and transfer method without dilution of glycerol. The technique being quite simple is worth trying in farm animals where importance of this technique in non-surgical transfer of frozen-thawed embryos will be a boon.

Animals↗
PubMed1989

A protocol used for splitting mouse embryos into two halves.

8-16 cell embryos and early blastocysts were obtained from the oviducts and anterior portion of uterine horns of albino mice at 70 and 90 hr after LH injection respectively. Splitting of embryos was done by using two microtools attached to a micromanipulator unit (Research Instruments Ltd, UK). After bisection, each pair of the half embryos is transferred to a dish containing 2 ml of T-6 medium and cultured in CO2 incubator (at 39 degrees C, 95% RH and 5% CO2 in air mixture). Splitting of blastocysts as compared to 8-16 cell embryos was found difficult (35.48% vs 52.44%, respectively). 38.88% of bisected 8-16 cell embryos and 11.36% of bisected blastocysts developed on 48 hr culture. Information on splitting mouse embryos and their subsequent development in culture are significant in view of using the technique for commercial application and for research in developmental biology of animal embryos.

Animals↗
PubMed1989

Cryopreservation of mouse embryos at -196 degrees C by vitrification.

Embryos (8-16 cell) were obtained from random bred albino mice (6-8 weeks old) that were induced to superovulate by injections of 5 I.U. PMSG and 5 I.U. hCG given 48 hr apart. Embryos were exposed to intracellular cryoprotecting medium (glycerol 10%, 1-2 propanediol 20% in PBS) for 10 min and then transferred to extracellular vitrification medium (25% glycerol, 25% 1-2 propanediol in PBS). Vitrification medium containing embryos, and diluent (1 M sucrose) were loaded in a straw and immediately plunged into liquid N2. After thawing at 20 degrees C, the contents of the straw were mixed by shaking (1 step dilution) and emptied in a petri dish. After 3 washings in culture medium the embryos were kept in CO2 incubator for further development. In 3-step dilution procedure the dilution of cryoprotectants was done in 0.5 and 0.25 M sucrose before culture. Embryos in 3-step dilution of cryoprotectants exhibited high survival as compared to 1-step dilution (20.23% vs 6.55%).

Animals↗
PubMed1985

How does embryo manipulation fit into present and future pig reproduction?

Available techniques for the collection and direct transplantation of pig embryos are simple and efficient and could be used for the expansion of new lines, for increasing selection pressure in nucleus herds and for extracting healthy stock from a diseased source. However, the reduced viability of pig embryos during culture in vitro and the inability as yet to preserve them by deep-freezing impose limits to the use of embryo transplantation for the export or import of potential breeding stock. The efficiency of breeding schemes could be improved by the sexing of embryos and the possibility of producing genetically identical twins or quadruplets by micromanipulation of embryos should improve the efficiency of animal experimentation. Chimaerism may be used to rescue embryos of a non-viable genotype such as parthenotes or those derived by hybridization, but the greatest revolution in pig breeding may be brought about by the introduction of foreign cloned genes into eggs and the production of transgenic animals. Eggs at an appropriate stage for microinjection may be provided in the future by techniques for the maturation and fertilization of oocytes in vitro. Animal breeders should be aware of the potential impact of techniques for the manipulation of eggs and embryos on future developments in animal production.

Animals↗
PubMed1978

The viability of deep-frozen cow embryos.

Day 7 cow embryos were frozen in 1.5 M-DMSO in PBS at 0.3 degrees C/min to -36 degrees C and at 0.1 degrees C/min between -36 and -60 degrees C before being plunged directly into liquid nitrogen. They were subsequently thawed (rapidly to -50 degrees C, at 4 degrees C/min from -50 to -10 degrees C, and rapidly again) to room temperature. Embryonic viability was tested by four different transfer techniques. Maximum pregnancy rate (8/12) was obtained with surgical transfer immediately after thawing and dilution of DMSO.

Animals↗
PubMed1978

A reciprocal translocation, t(6p+; 14q-), in the pig.

A reciprocal translocation, identified as t(6p+; 14q-), is described in a 38,XX intersex pig. It is the fourth reciprocal translocation to be reported for this species, whereas Robertsonian translocations, of frequent occurrence in cattle and sheep, are so far unknown in domestic pig breeds.

Animals↗
PubMed1976

The occurrence of C19 steroids in testicular tissue and submaxillary glands of intersex pigs in relation to morphological characteristics.

Five true hermaphrodite pigs and two male pseudohermaphrodite pigs were studied. A 38XX sex chromosome constitution was found in peripheral leucocytes of three true hermaphrodites and in one male pseudohermaphrodite; XX/XY mixoploidy was present in the leucocytes of the remaining male pseudohermaphrodite. The occurrence of C19 steroids, including 16-androstenes, in the testicular tissue and submaxillary gland of intersex pigs was of a similar pattern to that found previously in mature boars, and masculinization of the genital tract was related to the amount of testicular tissue present. It is postulated that in the absence of germ cells in the testicular tissue of intersex pigs the Sertoli cells may be involved in the metabolism of dehydroepiandrosterone to 5-androstenediol, a possible testosterone precursor in the pig. The high levels of 16-androstenes found in the submaxillary gland of intersex pigs indicates that these steroids are responsible for 'boar taint' in these animals. In contrast to the boar, no consistent relationship was found between the occurrence of C19 steroids and the degree of masculinization of the submaxillary gland; it is postulated that the predominantly female genetic constitution may have affected the response of the salivary gland to androgen.

Androstenediols↗
PubMed1976

Deep freezing of sheep embryos.

Sheep embryos, collected 1-8 days after oestrus, were placed in Dulbecco's phosphate-buffered saline medium (PBS). After treatment, the viability of the embryos was tested by temporary transfer to ligated rabbit oviducts. In Exp. 1, Days 5-8 embryos survived for at least 15 min at 0 degrees C in the presence of 1-5 M-DMSO. In Exp. 2, 12/14 Days 5-8 embryos survived after being frozen in 1-5 M-DMSO at 0-3 degrees C/min to temperatures ranging between-15 degrees and -60 degrees C and then thawed at 12 degrees C/min. In Exp. 3, Days 5-8 embryos were frozen in 1-5 M-DMSO at 0-3 degrees C/min to below-65 degrees C before being transferred to liquid nitrogen (-196 degrees C), and stored for 12 hr to 1 month. The embryos were thawed at 3 degrees C/min, 12 degrees C/MIN or 360 degrees C/min and, after transfer to rabbit oviducts, 0/4, 10/36 and 1/4, respectively, developed normally. The 11 embryos which were considered normal when recovered from the rabbit oviducts plus 1 slightly retarded embryo were transferred to 7 recipient ewes. Four ewes subsequently lambed, producing 5 lambs. In addition, 8 embryos were transferred to 4 ewes directly after thawing. Three of these ewes subsequently lambed, producing 3 lambs.

Animals↗
PubMed1976

Artificial insemination with spermatozoa in formaldehyde.

The ability of formaldehyde to preserve the integrity of the membranes of spermatozoa, as indicated by eosin staining (Dott & Foster, 1975), prompted an investigation to discover what other properties of spermatozoa were preserved by low concentrations of formaldehyde in vitro. In a series of experiments on bull, ram and boar spermatozoa it has been shown that spermatozoa rendered immotile by formaldehyde recovered their motility when the formaldehyde was removed by washing up to 12 hr afterwards (H.M. Dott & G.C. Foster, unpublished). To find out if fertility was preserved ewes and sows were inseminated with spermatozoa rendered immotile with formaldehyde.

Animals↗