Characteristics of an oviductal glycoprotein and its potential role in the fertilization process.
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Amino Acid Sequence↗
Biomedical subjects
J K Thibodeaux
Publications and source records attributed to J K Thibodeaux.
Enhancement of bovine oocyte fertilization in vitro with a bovine oviductal specific glycoprotein.
The purpose of this study was to determine the effect of a partially purified bovine oviductal glycoprotein (bOGP) on fertilization rates of bovine oocytes. The effect of albumin (control protein) or bOGP at 100 micrograms ml-1 during the 16-18 h fertilization period was evaluated in a standard IVF system using a sperm concentration between 0.5 and 0.125 x 10(6) spermatozoa ml-1. bOGP maintained a higher (P < 0.05) fertilization rate (62.0% versus 31.2%) at 0.125 x 10(6) spermatozoa ml-1 compared with the albumin control. The enhancement of fertilization by bOGP was blocked by the inclusion of a specific antibody to bOGP, whereas the antibody with albumin had no effect. A 2 h gamete preincubation step was subsequently included in the IVF procedure (0.125 x 10(6) spermatozoa ml-1) to determine whether the effect of bOGP was mediated through an interaction with the oocyte, the spermatozoon or both. When oocytes were preincubated with bOGP the fertilization rates were higher (P < 0.05) than with the albumin control (oocytes and spermatozoa exposed to albumin), whereas preincubation of spermatozoa with bOGP did not affect fertilization rates. There was no synergistic effect of preincubating oocytes and spermatozoa with bOGP. The increase in fertilization rate achieved by preincubating oocytes with bOGP was blocked with a specific antibody to bOGP. These results suggest that the increase in fertilization rates observed when bOGP is included during the 16-18 h fertilization period are primarily mediated through the interaction of bOGP with the oocyte since the same facilitatory effect was observed with a 2 h preincubation of oocytes before IVF. The ability to block these effects with a polyclonal antibody specifically generated against bOGP shows that this biological activity is due to bOGP. In summary, bOGP enhances fertilization in bovine oocytes whether it is included during preincubation or insemination and this appears to be due to a direct effect on the oocyte.
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Effects of dexamethasone administration to diestrus cows on systemic progesterone, estrogen and uterine cyclooxygenase production.
Parenteral administration of dexamethasone to diestrus cattle can extend the length of the natural estrous cycle. In mice, dexamethasone has been shown to inhibit production of the second isozyme of the cyclooxygenase (COX) enzyme (a rate limiting enzyme in prostaglandin formation). Therefore, the purpose of this study was to determine the effect of dexamethasone on estrous cycle length and COX-1 and -2 production by the uterine endometrium of cyclic cattle. Nine crossbred beef cows that exhibited two previous normal estrous cycles were randomly assigned to two treatments; a control group administered intramuscular injections of vehicle, and a dexamethasone group administered 8 mg of dexamethasone (Azium, Schering Corp., Kenilworth, NJ). Both groups received twice daily injections on day 13-22 of the treatment cycle. Uterine endometrial biopsies were collected on days 16, 19 and 22 of the treatment cycle. Blood samples were collected daily on day 13-22 of the treatment cycle for plasma progesterone and estradiol concentrations.
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Potential use of embryo coculture with human in vitro fertilization procedures.
PURPOSE: This review was designed to outline potential uses of an embryo co-culture system in human assisted reproduction programs to improve embryo quality and pregnancy rates. RESULTS: The various cell types used in embryo co-culture were reviewed in addition to the use of co-culture for both animal and human embryos. Co-culture provides a method to enhance embryo development in an inadequate in vitro environment without compromising embryo quality. Human IVF laboratories have used various types of "helper cells" to improve rate of development, reduce cell fragmentation rate and in some instances increases pregnancy and implantation rates. CONCLUSION: In conjunction with several assisted reproduction procedures such as IVF, microsurgical fertilization, cryopreservation and genetic evaluation, co-culture may increase the number of viable embryos for replacement and improve pregnancy rates.
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Effect of a serum extender containing growth factors on development of IVM and IVF bovine embryos.
The present experiments were conducted to determine if supplementation of the culture medium with a serum extender containing growth factors would increase development of bovine embryos into morulae or blastocysts, following in vitro maturation (IVM) and in vitro fertilization (IVF). In Experiment 1, bovine zygotes were cultured in CR1 medium supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 2, bovine zygotes were cultured in the presence of cumulus cells in CR1 medium supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 3, bovine oocytes were matured in Medium 199 supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 4, oocytes were matured in Medium 199 with 10% fetal bovine serum (FBS) or 5% FBS with serum extender. Following maturation, zygotes were cultured in CR1 medium with 10% FBS or 5 % FBS and serum extender. In all 4 experiments, the embryos were cultured in vitro until Day 7 after IVF, and development to the morula or blastocyst stage was assessed. The findings of the first 2 experiments showed that the serum extender did not directly influence embryo development but did stimulate development when cumulus cells were included in the culture system. The remaining 2 experiments showed that the serum extender did influence development through its interactions with cumulus cells during maturation and/or culture. These findings suggest that although growth factors or other products do not directly stimulate bovine embryo development their effects may be mediated through secondary cell systems.
Journal Article↗
Contact-associated interactions between large and small bovine luteal cells during the estrous cycle.
This experiment was designed to study the effects of cell-to-cell contact, arachidonic acid (10 microM; AA), oxytocin (10 microM), and luteinizing hormone (5 ng; LH) on bovine luteal cell function. Corpora lutea collected from Holstein cows between Days 10 and 12 (n = 4; midluteal stage) or 17 and 18 (n = 4; late-luteal stage) of the estrous cycle (Day 0 = estrus) were dispersed, and small and large cells were separated by unit gravity sedimentation and flow cytometry. Large and small luteal cells were either incubated together, allowing intercellular contact, or separately, without intercellular contact, with culture well inserts. Cells were incubated in a modified Ham's F-12-N-hydroxyethylpiperazine-N'-2-ethanesulfonic acid medium. After an 18-hr preincubation period, treatments were introduced and cells were incubated for 240 hr. Media samples were collected and treatments were replaced at 48-hr intervals. Incubations were maintained at 37 degrees C in 5% CO2 in humidified air. Overall, progesterone secretion decreased with increased incubation time (P < 0.0001), regardless of treatment, stage of the cycle, or cell arrangement. During the 18-hr pretreatment period, large and small luteal cells with contact secreted more progesterone than did luteal cells without contact during both the mid- (P < 0.0001) and late-luteal stages (P < 0.06) of the estrous cycle. After treatments were initiated, both mid- and late-stage luteal cells treated with LH secreted more (P < 0.0001) progesterone than occurred with any other treatment; oxytocin, AA, and control treatments were similar.(ABSTRACT TRUNCATED AT 250 WORDS)
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Interactions between large and small luteal cells collected during the mid- or late-luteal stages of the bovine oestrous cycle.
The effects of contact between large and small bovine luteal cells together with those of luteinizing hormone (LH) or arachidonic acid (AA) on progesterone production during the oestrous cycle were investigated. Corpora lutea were collected during the mid-luteal stage (Days 10-12; n = 4) and late-luteal stage (Days 17-18; n = 4) of the oestrous cycle. Large and small luteal cells were dispersed and separated and then incubated together or separately. Mid-luteal stage cells were treated with LH (0 or 5 ng) whereas late-luteal stage cells were treated with LH (0 or 5 ng) or AA (0 or 10 microM). Culture medium was collected and replaced 1, 3 and 6 h after starting treatments. Progesterone production decreased (P < 0.0001) with increased incubation time irrespective of cell arrangement, the stage of the oestrous cycle or treatment. During the 18 h before treatment, cells in the contact arrangement produced more progesterone (P < 0.003) than cells without contact in both mid- and late-luteal stages of the oestrous cycle; progesterone production within cell arrangements between prospective treatment groups was similar. After initiating treatments, mid-luteal stage cells in the control group without contact produced more progesterone (P < 0.01) than cells with contact. Mid-luteal stage cells treated with LH produced more (P < 0.0001) than control cells; progesterone production between cell arrangements within the LH treatment group was similar. In the late-luteal stage cells, both LH and AA increased (P < 0.01) progesterone production by comparison with control cells; LH and AA treatment groups produced similar results.(ABSTRACT TRUNCATED AT 250 WORDS)
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Effect of media substitutes on bovine granulosa cell function and proliferation during in vitro culture.
The development of a serum-free culture system for bovine granulosa cells that would allow for cellular proliferation without induction of steroidogenesis would provide researchers with an important in vitro tool for determining differentiation mechanisms during folliculogenesis. The objective of the present study was to determine the effect of a commercially prepared serum substitute and a medium supplement on proliferation and progesterone production by bovine granulosa cells. Granulosa cells were obtained by aspirating the follicular fluid of follicles 2 to 8 mm in diameter. For each experiment, growth curves to determine the proliferative and steroidogenic response of granulosa cells to several different medium additions were constructed. Cells were counted on d 1, 2, 4, 6, and 8 of culture to determine cell concentration and the media harvested to determine progesterone content. In Exp. 1, granulosa cells were seeded at an initial rate of 5.0 x 10(5) for 48 h in serum-supplemented medium then allotted to one of five treatments including medium alone or medium containing fetal bovine serum (FBS; 1%), Gibco BRL media supplement-x (GMS-X; 1%), fatty acid-free bovine serum albumin (FAF-BSA; 4 mg/mL), or SerXtend (5%). For Exp. 2 and 3, granulosa cells were plated in serum-supplemented medium for either 48 or 24 h and seeded at either 5.0 x 10(5) or 2.5 x 10(5) cells/flask, respectively, and cultured in different concentrations of SerXtend. All treatment media supported granulosa cell proliferation to some extent; the SerXtend treatment provided the highest proliferation rate at all concentrations above .3125%. Also, during the proliferative stage of the growth curve, cells in the SerXtend treatment produced lower amounts of progesterone compared with cells in the other treatments. In summary, granulosa cells may be propagated in vitro in a serum-free environment without inducing progesterone production.
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Established cell lines and their conditioned media support bovine embryo development during in-vitro culture.
These experiments were conducted to evaluate the ability of different somatic-cell monolayers or conditioned medium from somatic cells for supporting bovine embryo development in vitro. In the first experiment, bovine embryos (2- to 4-cells) were allocated randomly to a control (medium 199 with 10% fetal bovine serum and antibiotics) group or co-cultured with bovine oviduct epithelial (BOEC), buffalo rat liver (BRL), Madin Darby bovine kidney (MDBK) or African green monkey kidney (Vero) cells. In the second experiment, bovine embryos (1-cell) were allocated randomly to the following groups: control medium or conditioned medium from BOEC, BRL, MDBK and Vero monolayers. In both experiments, development to the blastocyst stage was assessed after 8 days of incubation at 39 degrees C and 5% CO2. In Experiment 1, coculture improved development to the blastocyst stage compared with control medium alone, and the highest development was observed after co-culture with BOEC. In Experiment 2, conditioned medium enhanced development to morulae and blastocysts compared with the control medium; however, no differences were detected among different cell supports. These results indicate that both co-culture and conditioned medium from different cell monolayers supported development to the blastocyst stage at a higher efficiency than control medium alone.
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Stimulation of progesterone production in bovine luteal cells by co-incubation with bovine blastocyst-stage embryos or trophoblastic vesicles.
A study was conducted to determine whether bovine blastocyst-stage embryos and trophoblastic vesicles stimulate the production of progesterone in bovine luteal cells during incubation in vitro. The effects of co-incubation of these embryos and vesicles with uterine endometrial tissue on progesterone production was also investigated. Bovine small and large luteal cells were obtained on day 12 of the oestrous cycle, dispersed by unit gravity sedimentation and recombined to provide preparations free of accessory cells. Blastocyst-stage embryos were obtained on day 7 and trophoblastic vesicles were obtained from bovine embryos on day 12. A uterine endometrial tissue sample was obtained from the same cow from which the corpus luteum was taken. Treatment groups were arranged in 24-well plates as follows: luteal cells alone; luteal cells and one trophoblastic vesicle; luteal cells and one blastocyst embryo; luteal cells and a 10 mg uterine endometrial sample; luteal cells, one trophoblastic vesicle and a uterine endometrial sample; and luteal cells, one blastocyst embryo and a uterine endometrial sample. All treatment groups were incubated (at 37 degrees C under 5% CO2) in Ham's F-12 medium supplemented with antibiotics (100 micrograms penicillin ml-1 and 100 U streptomycin ml-1, L-glutamine (0.29 mg ml-1), insulin (5 micrograms ml-1), transferrin (5 micrograms ml-1) and selenium (5 ng ml-1) for 12 h. Samples of the medium were harvested 10 min (basal concentration) and 2, 6 and 12 h after incubation to determine the concentrations of progesterone and prostaglandin.(ABSTRACT TRUNCATED AT 250 WORDS)
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Interactions between large and small bovine luteal cells in a sequential perifusion co-culture system.
The objectives of this experiment were to study large and small luteal cell interactions and examine the effect of arachidonic acid (AA) on progesterone production by separated bovine luteal cells. Corpora lutea collected from Holstein cows (n = 5) on d 12 of the estrous cycle were dispersed, and small (SLC) and large (LLC) luteal cells were separated by unit gravity sedimentation and flow cytometry. Cells were incubated at 37 degrees C in separate perifusion chambers with a modified Ham's F-12-HEPES medium and aerated with 95% O2:5% CO2. The flow rate of medium was 100 microL/min, and fractions were collected at 30-min intervals for 4 h. Luteal cells were arranged in tandem so that medium from the first cell type would pass through the chamber containing the second cell type. Luteal cells were arranged so that medium flowed from 1) SLC to SLC, 2) LLC to LLC, 3) SLC to LLC, 4) LLC to SLC, 5) SLC to LLC, 6) LLC to SLC; medium for arrangements 5 and 6 contained 10 microM AA. Cells in arrangements 5 and 6 were perifused for 30 min before AA was added. Progesterone was measured with an enzymeimmunoassay. The LLC to LLC arrangement had a greater (P < .05) average progesterone secretion rate than all other cell arrangements, and the SLC to SLC arrangement had the least progesterone secretion rate.(ABSTRACT TRUNCATED AT 250 WORDS)
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Frozen-thawed cumulus-granulosa cells support bovine embryo development during coculture.
OBJECTIVE: To evaluate the ability of bovine cumulus-granulosa cells to survive cryopreservation and subsequently support bovine embryo development during coculture. DESIGN: In vitro-matured and -fertilized bovine embryos (two- to four-cell) were allotted randomly to one of three treatment groups: [1] control medium alone consisting of Medium 199 containing 10% fetal bovine serum and antibiotics, [2] cocultured on fresh bovine cumulus-granulosa cells in control medium, or [3] cocultured on frozen-thawed cumulus-granulosa cells in control medium. Embryo development was assessed on days 7 and 8 after IVF. RESULTS: Coculture improved embryo development on days 7 and 8 compared with the control group. However, embryo development on days 7 and 8 did not differ among coculture groups. CONCLUSIONS: Frozen-thawed cumulus-granulosa cells enhance embryo development similar to fresh cells during in vitro coculture.
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The effects of different types of syringes on equine spermatozoa.
Control extender was incubated at 4 degrees C for 24 hours. Rubber or plastic syringe plungers were separately incubated in semen extender for 24 hours at 4 degrees C. Following incubation, the extender was stored at -20 degrees C until the time of semen collection. The treatments consisted of the following: Group A = equine semen plus control extender; Group B=equine semen plus extender incubated with rubber plungers and Group C=equine semen plus extender incubated in plastic plungers; Group D=equine semen plus control extended in rubber plunger syringes and Group E=equine semen plus control extender in plastic plunger syringer. Each group contained a 5-ml volume of semen and extender at a concentration of 1.0 x 10(8) sperm/ml. The number of live spermatozoa, percentage of progressively motile spermatozoa and rate of progressive motility were taken following collection and every 15 minutes for 1 hour following application of treatments. In experiment 2, treatments were allowed to incubate with semen for 45 minutes, then the extender was removed and was replaced with fresh extender. The rate of progressive motility and the percentage of progressively motile spermatozoa were taken immediately, at 45 minutes, and then every 15 minutes for 1 hour. In experiment 1, the number of live spermatozoa was not affected among the 5 groups. However, there was a decrease (P<0.01) in the rate of progressive motility and in the percentage of progressively motile spermatozoa in Group B compared with the remaining 4 treatment groups at 30, 45 and 60 minutes, with no differences noted when semen was held in syringes with a rubber or a plastic plunger. In experiment 2, the percentage of progressively motile spermatozoa increased after the addition of the control extender.
Journal Article↗
Role of platelet-derived growth factor in development of in vitro matured and in vitro fertilized bovine embryos.
This experiment was designed to determine whether the stimulatory effects of bovine oviductal epithelial cells (BOEC) on development of early bovine embryos are due to platelet-derived growth factor (PDGF). Four hundred and twenty five 8-cell bovine embryos derived from in vitro maturation and in vitro fertilization procedures were equally and randomly allotted to one of the following culture treatment groups: control medium alone (Menezo's B2 medium; MB2), MB2 with 1 ng PDGF ml-1 (PDGF), 1 ng PDGF ml-1 and 10 micrograms anti-PDGF antibody ml-1 (PDGF + Ab), BOEC or BOEC and 10 micrograms anti-PDGF antibody ml-1 (BOEC + Ab). All embryos were cultured in 100 microliters of serum-free MB2 medium supplemented with 2 mg fatty-acid-free bovine serum albumin ml-1. Embryos for all treatment groups were incubated at 39 degrees C and 5% CO2 in humidified air in groups of five embryos per well in 96-well culture plates until 7 days after in vitro insemination. A higher proportion of embryos developed to > 8-cell and to the morula stage following culture with PDGF, BOEC or BOEC+Ab than with MB2 alone. Incubation of PDGF and BOEC-treated embryos with anti-PDGF reduced development to the morula and blastocyst stages. However, anti-PDGF did not completely inhibit blastocyst development when added to BOEC. In addition, embryos incubated with BOEC and anti-PDGF contained a reduced number of inner cell mass cells compared with embryos incubated with BOEC alone. These results indicate that PDGF provides a developmental stimulus similar to BOEC for bovine embryos at the fourth cell cycle.(ABSTRACT TRUNCATED AT 250 WORDS)
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Stimulation of development of in vitro-matured and in vitro-fertilized bovine embryos by platelets.
In vitro-fertilized bovine embryos were incubated in Menezo's B2 medium (MB2) supplemented with 2 mg/mL of BSA. In Exp. 1, eight-cell stage embryos were allotted to one of the following groups: control medium (MB2), MB2 with 20 ng/mL of platelet-activating factor (PAF), 1 x 10(7) bovine blood platelets (Platelets), oviductal cells (BOEC), BOEC and 20 ng/mL of PAF (BOEC+PAF), or BOEC and 1 x 10(7) platelets (BOEC+Platelets). In Exp. 2, eight-cell embryos were allotted to one of the following groups: control medium (MB2), MB2 with 1 x 10(7) platelets (Platelets), 1 x 10(7) platelets and 10 micrograms/mL of platelet-derived growth factor antibody (Platelets+anti-PDGF), 1 x 10(7) platelets and 1 microgram/mL of indomethacin (Platelets+Indomethacin), or 1 x 10(7) platelets and 3 micrograms/mL of mianserin (Platelets+Mianserin). Embryos were incubated at 39 degrees C in 5% CO2 in groups of five until 8 d after in vitro fertilization (IVF). In Exp. 1, Platelets stimulated embryo development to the morula, blastocyst, and expanded blastocyst stages. Embryo development was greatest in the BOEC+Platelets group on d 7 and 8 after IVF. Only embryos incubated in the BOEC+Platelets treatment group reached the hatched blastocyst stage on d 8. In Exp. 2, embryos incubated in the Platelets treatment group had the greatest (P < .05) proportion develop beyond the eight-cell stage. Embryos incubated in the Platelets + anti-PDGF group had less (P < .05) development beyond the eight-cell stage and to the morula stage. These results indicate that the stimulatory effects of PDGF on bovine embryo development may be derived from both the oviductal epithelium and platelets.
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Intrauterine infusion of prostaglandin E2 and subsequent luteal function in cattle.
The objective was to evaluate the effect of intrauterine infusion of prostaglandin E2 (PGE2) on luteal function in cattle. Heifers and cows were randomly assigned after two normal estrous cycles to either PGE2 or control treatment groups. Females in Treatment A were infused with 1 mg of PGE2 once daily into the uterine horn ipsilateral to the corpus luteum between days 7-10 of the estrous cycle with a 0.25 ml plastic semen straw and an artificial insemination pipette. Females in Treatment B were similarly infused with 1 mg of PGE2 once daily in 20 ml of a carrier vehicle via a catheter on days 10 and 11 of the estrous cycle. Control animals were infused with the carrier vehicle using either a semen straw (Treatment C) or via a catheter (Treatment D) on the same days of the estrous cycle. Blood samples were collected daily to monitor plasma progesterone concentrations during the treatment period. Females infused with PGE2 on days 7-10 of the estrous cycle returned to estrus in a mean of 23.5 days (range 22-25 days) and were similar (P > 0.05) to those infused on days 10 and 11 which returned to estrus in 23.5 days (range 22-25 days). Animals similarly infused with carrier vehicle on the same days of the estrous cycle returned to standing estrus in 20.2 days (range 17-23 days). Plasma progesterone concentrations indicated an extended period of elevated progesterone concentrations in PGE2-treated animals compared with control animals. These results indicate that short term administration of PGE2 early in the estrous cycle may result in extended luteal maintenance.
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Evaluating an in vitro culture system of bovine uterine and oviduct epithelial cells for subsequent embryo co-culture.
Three experiments were conducted to evaluate the effects of culture medium and incubation temperature on bovine uterine and oviduct epithelial cell growth, so that the most efficient combination could then be used to develop a co-culture system for bovine embryos. In the first experiment, uterine and oviduct epithelial cells at either the second or third subpassage were incubated for 8 days at 37 degrees C with 5% CO2 in Tissue Culture Medium-199, CMRL-1066, Minimal Essential Medium, Ménézo's B2 or Ham's F-12 medium. In addition to plotting growth curves of cell populations, the cell cycle was monitored for 8 days by flow cytometry. Uterine and oviduct epithelial cells incubated in CMRL-1066 exhibited the highest growth rates during the 8-day culture period. However, there were no differences in cell cycle analysis among treatment groups during the incubation period. In the second experiment, CMRL-1066 medium was used to evaluate growth and proliferation of uterine and oviduct epithelial cells incubated at 37 degrees C or 39 degrees C; temperature had no significant effect on growth rates or proliferation rates for either uterine or oviduct cells during the 8-day incubation. In the third experiment, the more promising culture media for epithelial cell culture studies were chosen for in vitro maturation and subsequent in vitro fertilization (IVF) of bovine oocytes. Early cleavage-stage embryos produced by IVF procedures were subsequently cultured in vitro for 7 days in medium alone or with oviduct epithelial cells. In this study, the culture medium did not influence fertilization or cleavage rates. However, more embryos co-cultured with oviduct epithelial cells were considered viable after 7 days of incubation compared with embryos incubated in medium alone. These results indicate that various incubation conditions can influence the growth of bovine uterine and oviduct epithelial cells in vitro. However, in spite of changes in cell growth patterns, there does not appear to be a change in their embryotropic capabilities in vitro.
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In-vitro co-culture of early stage caprine embryos with oviduct and uterine epithelial cells.
Early stage caprine embryos were incubated with goat oviduct and uterine cells to evaluate whether these cells could be used as a somatic cell culture system to enhance development through the developmental block at the 8- to 16-cell stage during in-vitro culture. Following gonadotrophin treatment and natural mating, 2- to 4-cell embryos were surgically recovered from donor females for in-vitro culture studies. In Experiment 1, embryos were equally and randomly allotted to culture treatments of either culture medium plus caprine oviduct cells or culture medium alone. In both treatment groups, embryos were incubated in Medium-199 with 10% fetal bovine serum, 0.25% lactalbumin and 1% antibiotic-antimycotic at 37 degrees C in a humidified atmosphere of 5% CO2 in air. In Experiment 2, similar embryos were cultured in the same medium with either caprine oviduct cells, caprine uterine cells or sequentially incubated with oviduct cells and then uterine cells during a corresponding incubation interval. The culture conditions in Experiment 2 were the same as in Experiment 1. Following 72 h in culture, (Experiment 1), significantly more embryos developed through the in-vitro developmental block into blastocysts and hatched blastocysts when cultured with oviduct cells compared with no embryos developing through the in-vitro block when incubated with medium alone. In Experiment 2, caprine embryos co-cultured with oviduct cells alone resulted in more embryos developing into blastocysts and hatched blastocysts compared with those co-cultured with uterine cells alone.(ABSTRACT TRUNCATED AT 250 WORDS)
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