Biomedical subjects

J K Thibodeaux

Publications and source records attributed to J K Thibodeaux.

26 records · Page 2Linked to original sources

PubMed1992

Coculture of in vitro fertilized bovine embryos with oviductal epithelial cells originating from different stages of the estrous cycle.

Bovine embryos derived from in vitro fertilization procedures were cocultured in vitro with oviductal cells obtained from heifers between d 4 and 6 or d 14 and 16 of the estrous cycle. In addition, proteins secreted by oviductal cells isolated between d 4 and 6 or d 14 and 16 of the cycle were monitored. Embryos (2- to 4-cell) were incubated in Tissue Culture Medium-199 with 10% fetal bovine serum with or without oviductal cells at 39 degrees C for 10 d following in vitro insemination. There were more morulae, blastocysts, and hatched blastocysts following coculture with oviductal cells than with culture in medium alone. However, no differences were noted in embryo development following coculture with oviductal cells obtained between d 4 and 6 or d 14 and 16 of the estrous cycle. Also, no differences were detected in the amount of [35S]methionine-labeled proteins secreted by oviductal cells isolated from different days of the estrous cycle. These results indicate that oviductal epithelial cells isolated from early and late luteal phases of the estrous cycle will effectively support early embryonic development following prolonged in vitro culture.

Animals↗

PubMed1992

Effect of feeding various levels of sodium zeolite A on milk yield, milk composition and blood profiles in thermally stressed Holstein cows.

Mid-lactation Holstein cows (n = 48) were equally and randomly assigned to one of four feeding treatments of sodium zeolite-A (SZA). SZA was mixed in a grain mixture (50:50 grain to forage ratio) of 0% (control), 0.5%, 1.0% and 1.5% SZA on a dry matter intake basis. Cows were fed alfalfa hay in the first phase and corn silage in the second phase of the study as roughage sources. Milk samples were taken three times weekly (am and pm) and analyzed for milk fat, protein and lactose with blood profiles conducted from samples collected weekly. SZA significantly (P less than .05) increased feed intake at all three levels for both diets. Milk yield was significantly (P less than .05) greater in the alfalfa diet. However, milk fat percent and percent protein were greater (P less than .05) in the corn silage diet. The addition of SZA to the corn silage diet increased (P less than .05) milk fat percent at the 1.0% level and milk protein at the 1.5% level. Calcium in milk was significantly (P less than .01) increased and respiration rates significantly lowered (P less than .05) in both diets at the 1.0% level. Serum calcium was higher (P less than .05) at the 1.0 and 1.5% level in the hay diet and the 1.5% level in the corn silage diet. Also, serum glucose and alkaline phosphate levels were significantly (P less than .05) higher in the corn silage diet.

Aluminum Silicates↗

PubMed1992

In vitro enhancement of early-stage embryos with co-culture.

Over the years, a great deal of effort has been made to maintain the viability of mammalian embryos once they have been removed from the female's uterus. Early embryo research studies used the simplest of medium (eg, physiological saline) in an effort to maintain embryo homeostasis in vitro, but little progress on embryo viability was made. The addition of heat-treated serum, buffers, and essential amino acids to culture media have shown evidence that improvements could be made in culturing mammalian embryos in vitro; however, it has become clearly evident that this was not going to be a simple task. In recent years, the most notable progress made in this research area has been made with the use of "helper" cells to co-culture early-stage embryos to hatched blastocysts. This article reviews the classic works that have contributed to the development of embryo culture systems that are now used in genetic engineering research and commercial embryo transplant units in the livestock industry. The development of trophoblastic vesicle culture methods and uterine cell co-culture systems were basic contributors to the development of the mammalian embryo co-culture systems as we know them today. Recent studies with follicular granulosa cells and oviduct epithelial cells have added much to our understanding of embryo development in vitro. Novel co-culture systems, such as culturing mammalian embryos in the amnion of a developing chick embryo, certainly offer encouragement that research efforts should continue until an optimal culture system is developed for each mammalian species. The potential uses of embryo culture systems for both humans and animals have yet to be fully understood.

Animals↗

PubMed1992

The use of image analysis to evaluate the development of uterine and oviduct epithelial cells during in vitro culture. A potential quality assurance procedure for in vitro laboratories.

The objective of this study was to establish selection criteria for morphologic assessment of cell quality using a computer image-analysis system. Uterine and oviduct epithelial cells were isolated from the reproductive tracts of cyclic cows using a trypsin solution. Harvested cells were cultured in Tissue Culture Medium-199 with 10% fetal bovine serum and maintained at 37 degrees C with 5% carbon dioxide in air. Subcultures of different confluent monolayers were established using a weak trypsin solution. Morphologic assessment of cell integrity and viability were made visually, with electron microscopy and image analysis. After morphologic assessment, cells were assigned quality scores. Evaluations were conducted on cells during primary culture and following the first and third subpassages. Image-analysis evaluation was conducted on cells using a silicon-intensified target camera and computer-based software. Gray-level density was determined on cells from each flask. There was no difference (P greater than .05) between sample replicates, indicating repeatable measurements were obtained with the image-analysis system. In addition, there was no significant difference in parameters measured between uterine and oviduct cells; therefore, data from both cell types were combined for further statistical comparisons. Gray-level density values for combined uterine and oviduct cells during primary culture and following the first and third subpassages were as follows: 139.7, 154.5, and 173.3, respectively. There was an increase (P less than .05) in gray-level density values with each subpassage of the cell populations. Furthermore, gray level was influenced (P less than .01) by cell quality for combined uterine and oviduct cell populations. These results indicate that epithelial cell quality during in vitro culture may be effectively determined using image analysis. This approach should not be overlooked in establishing quality control measures for uterine and oviduct cells before mammalian embryos are cocultured in vitro.

Animals↗

PubMed1991

Effects of stage of the bovine oestrous cycle on in-vitro characteristics of uterine and oviductal epithelial cells.

The objective of this experiment was to evaluate the effects of stage of the bovine oestrous cycle on in-vitro morphology, growth and monolayer foundation of uterine and oviductal epithelial cells. Epithelial cells were isolated from the uterus and oviducts collected from cyclic cattle on the day of oestrus (Treatment A), and between days 4 to 6 (Treatment B), days 8 to 10 (Treatment C) and days 14 to 16 (Treatment D) of the oestrous cycle. The morphological development, per cent cell viability and cell attachment were evaluated during primary culture and after the first and third subpassages. The highest per cent cell viability and cell attachment during primary culture, respectively, were noted in Treatment B for both uterine (87.7 and 87.5%) and oviductal (88.4 and 87.2%) cell populations. Uterine epithelial cell populations in Treatments C and D, respectively, had the lowest viability (76.5 and 68.8%) and attachment (10.8 and 10.5%) during primary culture. There were marked improvements in cell viability and cell attachment following the first subpassage (P less than 0.001) compared with primary cultures for both uterine and oviductal cells. These results indicate that the stage of the oestrous cycle has dramatic effects on uterine and oviductal epithelial cell morphology and developmental patterns during primary in-vitro cultures. The stage of the oestrous cycle when cells are collected may be more important than was once realized when culturing early stage embryos in vitro.

Animals↗

PubMed1990

Early pregnancy associated thrombocytopenia as an initial response to pregnancy in cattle.

This study was conducted to determine if early pregnancy-associated thrombocytopenia exists in cattle as has been demonstrated in mice and in humans. Three experiments were designed to compare peripheral platelet counts in pregnant versus nonpregnant animals. In Experiment 1 heifers (n = 25) were artificially inseminated 12 h after the onset of estrus. Peripheral platelet counts in 19 pregnant versus 6 nonpregnant heifers did not reveal any significant differences between groups after insemination. In Experiment 2 embryos were collected nonsurgically from superovulated cows (n =18) on Days 6 to 7 after estrus. Platelet counts were monitored every 12 h after the first insemination until 60 h after the second insemination. Platelet counts and the number of embryos collected nonsurgically from these superovulated donors did not show any significant correlations (P>0.05). Ten recipient heifers synchronized to donor animals received either an unfertilized ovum or a good quality embryo via nonsurgical transfer into the uterus. There were no significant reductions in platelet counts after transfer. In Experiment 3 platelet counts were monitored daily in four pregnant and five nonpregnant recipient heifers between Day 0 and Day 30 after embryo transfer on Day 8 of the cycle. The platelet counts did not reveal any significant differences between the pregnant and nonpregnant groups throughout Days 0 to 30. These results indicate that early pregnancy-associated thrombocytopenia cannot be demonstrated in cattle. Peripheral platelet counts cannot be used as an indicator of early pregnancy in cattle.

Journal Article↗

PubMed1990

The refractometer index as a correction factor for urinary estradiol in rhesus females.

Refractometer indexes were standardized and found to be highly correlated (r = 0.831) to actual creatinine levels. Urinary estradiol corrected by creatinine levels and refractometer indexes were found to be highly correlated (r = 0.857). Predicted ovulation was the same day in 86% of the ovulatory cycles predicted by creatinine and refractometer corrected estradiol levels. Refractometer indexes may be used in place of creatinine levels to correct for urine concentration fluctuations when predicting ovulation in the rhesus female.

Animals↗

PubMed1989

An efficient procedure for manual platelet counting.

Blood samples were collected by jugular venipuncture from 15 dairy heifers and the blood platelets were counted by manual methods. The platelets were found to be uniformly distributed across the rows, columns and sides of a Neubauer haemocytometer, and it was shown that counting any 10 squares on either side of the haemocytometer and multiplying by a constant factor would accurately predict the total platelet count. This procedure would greatly reduce the time required to count large numbers of platelets per sample, and reduce errors due to the fatigue associated with counting large numbers of samples.

Animals↗