Biomedical subjects

R H Foote

Publications and source records attributed to R H Foote.

At least 55 records · Page 3Linked to original sources

PubMed1995

Effect of inositol and glycine with increasing sodium chloride and constant osmolality on development of rabbit embryos.

PURPOSE: Many commercially available culture media have a high sodium content as sodium chloride and sodium bicarbonate. We examined the effects of osmolytes, inositol and glycine, on embryos cultured in synthetic RD medium with a lower concentration of NaCl (93 mM) and a higher concentration of NaCl (116 mM) with media held constant at 270 mosmols. RESULTS: There were no significant effects of either glycine or inositol on embryo growth when the embryos were cultured in RD medium with 93 mM NaCl (P > 0.05). Culture of one-cell embryos for 72 h in RD medium with 93 mM NaCl, 116 mM NaCl and 116 mM NaCl containing 0.56 mM inositol resulted in 77%, 14% and 55% expanded blastocysts, respectively (P < 0.05). Corresponding values for two-cell embryos cultured for 67 h were 84%, 34% and 66% expanded blastocysts (P < 0.05). When one-cell and two-cell embryos were cultured in RD medium with 93 mM NaCl, 116 mM NaCl, 116 mM NaCl plus 1 mM glycine and 116 mM NaCl plus 1 mM glycine and 0.56 mM inositol, expanded blastocysts from one-cell embryos were 79%, 19%, 49%, and 48%, respectively (P < 0.05), and expanded blastocysts from two-cell embryos were 91%, 32%, 52%, and 53%, respectively (P < 0.05). CONCLUSION: Inositol and glycine presumably behave as osmolytes in providing substantial protection for rabbit one-cell and two-cell embryos cultured in a medium with high NaCl concentration.

Animals↗

PubMed1995

Pulsatile release of FSH for superovulation in cattle.

The studies reported here were directed towards the development of an implantable microcapsule which "pulses" release of follicle stimulating hormone, FSH, for application to superovulating cows. Final dose forms were administered using membrane-coated cylinders. The "pulse" of the FSH is achieved by membrane encapsulation of an effervescent/swelling core containing citric acid, sodium bicarbonate, glucose and FSH. Entry of water results in sufficient pressure increase (by gas generation) to rupture ("burst") the membrane. Time to rupture is dependent upon several factors, such as membrane permeability and thickness, and core composition and loading. The final dose forms were implanted by means of a trochar. This system was tested in sheep to substantiate in vivo "burst" times and then tested in cows to determine efficacy. In vivo burst times in sheep varied from 8 to 96 hr, based upon maximal FSH values in blood serum, and generally paralled the planned times resulting from in vitro tests. Multiple capsules designed to release FSH as a pulse or steady state were tested on a limited number of cows plus a control (n = 10). Four of the combinations resulted in 11, 11, 14 and 16 ovulations, indicating that further development has promise of providing a one-injection system using FSH for superovulating cattle.

Journal Article↗

PubMed1995

Development of early bovine embryos in co-culture with KSOM and taurine, superoxide dismutase or insulin.

Three experiments, utilizing 2578 embryos, were designed to test the effects of media, taurine, Superoxide dismutase and insulin on the development of embryos produced by in vitro maturation and in vitro fertilization (IVM/IVF). Embryos showing at least 1 cleavage during culture for 40 to 44 h after IVM/IVF were selected for further culture under various conditions for 6 d at 39 degrees C in 5% C0(2):95% air. A Buffalo rat liver (BRL) cell co-culture was used in all 3 experiments. Experiment 1 was a 3 x 2 factorial arrangement with KSOM (a high potassium simplex optimization-derived medium containing only 12 ingredients), Menezo B(2) and TCM-199 media with or without 7 mM taurine. Blastocyst production in the 3 media, respectively, was 48, 36 and 29% (P<0.05). Addition of 7 mM taurine increased the percentage of blastocysts from 34 to 42 (P<0.05). In Experiment 2, Superoxide dismutase (SOD) did not improve blastocyst development (P>0.05). In Experiment 3, insulin (75 ng/ml) added to KSOM resulted in 46% morulae plus blastocysts compared with 35% for the control (P<0.05). These results indicate that the co-culture of embryos in KSOM with taurine or insulin added is superior to commonly used complex media for efficient production of blastocysts following IVM/IVF.

Journal Article↗

PubMed1995

Effects of amino acids on the development of in-vitro matured/in-vitro fertilization bovine embryos in a simple protein-free medium.

The effects of commercially available amino acids, referred to as Eagle's non-essential amino acids (NEAA) and essential amino acids (EAA), on the development of bovine embryos produced by in-vitro maturation and in-vitro fertilization (IVF) in a simple modified protein-free KSOM medium containing polyvinyl alcohol (PVA) were studied. Embryos produced by IVF were cultured in an atmosphere of 5% O2, 5% CO2 and 90% N2 at 39 degrees C for the first 40-44 h in modified KSOM. Then embryos with four or more cells were cultured in modified KSOM-PVA with different amino acids for another 6 days in experiments 1 and 2. In experiment 3, to study embryo hatching and degeneration further as criteria of medium suitability, the culture time was 8 days. In experiment 1, more blastocysts were produced (P < 0.05) in the medium with 0.5 x NEAA (37%) than with 0.25 or 0.5 x EAA (both 15%). In experiment 2, when 0.0, 0.5 and 1.0 x concentrations of NEAA were combined with 0.0 and 0.5 x concentrations of EAA, the combination 1.0 x NEAA with 0.5 x EAA resulted in the most blastocysts (39%), with 11% hatching (P < 0.05). Without NEAA, 1.0 x EAA resulted in 6% blastocysts (P < 0.05). In experiment 3, KSOM-PVA with 1.0 x NEAA plus 0.5 x EAA resulted in 41% blastocysts, similar to the 40% blastocysts in KSOM-bovine serum albumin (BSA) with the same amino acids added. When culture was continued for 2 more days, more embryos hatched in these two treatments. These results indicate that the combination of 1.0 x NEAA with 0.5 x EAA promoted blastocyst formation and hatching, but EAA alone was detrimental. With PVA successfully replacing BSA, a completely defined medium resulted, which was equal or superior to complex medium, and is potentially useful for the culture of embryos from other species, such as humans.

Amino Acids↗

PubMed1995

Development of bovine embryos in KSOM with added superoxide dismutase and taurine and with five and twenty percent O2.

To further define requirements for embryo development without the assistance of complex media, serum, or coculture, the effects of 5% and 20% O2 concentrations, superoxide dismutase (SOD) (experiment 1), and taurine (experiment 2) were tested in a single culture medium (KSOM modified with 2.5 mM HEPES). Bovine embryos were produced in vitro maturation (IVM) and in vitro fertilization (IVF) of oocytes. After fertilization, embryos were cultured in 5% O2:5% CO2:90% N2 at 30 degrees C for the first 40-44 h; then embryos with 4 to 8 cells were cultured in different treatments for another 6 days. Percentage development subsequently was based on this number. In experiment 1, more blastocysts were produced in 5% O2:5% CO2:90% N2 (37%) than in 20% O2:5% CO2:75% N2 (18%; p < 0.01). Blastocyst development with 0, 300, and 600 U/ml of SOD, respectively, was 26%, 26%, and 30% (p > 0.05). In experiment 2, more blastocysts were formed in 5% O2 (39%) than in 95% air (30%; p < 0.05). Also, with 20% O2 and 0, 7, and 14 mM taurine in the medium, blastocyst formation was 18%, 35%, and 36% (p < 0.05). However, in 5% O2, 7 and 14 mM taurine was not beneficial (p = 0.99), as 39% blastocysts were formed with no taurine. In conclusion, results from both experiments demonstrated that 5% O2 was superior to 20% O2 for development of IVM/IVF bovine embryos. SOD had no effect in either 5% or 20% O2. However, 7 mM and 14 mM taurine counteracted the negative effect of 20% O2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Beneficial effects of culturing rabbit zygotes to blastocysts in 5% oxygen and 10% carbon dioxide.

Zygotes were collected from superovulated Dutch-belted rabbits 19 h after injection of LH and insemination. Oocytes that appeared to be unfertilized were discarded. The zygotes were distributed equally within each donor female across all culture treatments. Culture dishes contained 500 microliters of macromolecule-free RD medium consisting of equal parts of RPMI 1640 and low glucose Dulbecco's modified Eagle's medium. Embryos were cultured at 39 degrees C in several gas combinations of N2 plus the following: (1) 1% O2:10% CO2; (2) 5% O2:10% CO2; and (3) 20% O2:10% CO2. The control (4) was 95% air:5% CO2. The experiment was replicated with embryos from 11 donors providing 295 usable zygotes. After 84 h of culture, the percentages of blastocysts formed in treatments 1 to 4, respectively, were 13, 86, 82 and 59 (P < 0.01). The corresponding mean cell counts, including all cleaved embryos cultured (but not degenerate ones), were 55, 183, 118 and 68 (P < 0.01). These results indicate that 10% CO2 combined with 5% O2 is a more effective gas phase for culturing rabbit zygotes in a synthetic medium than is the commonly used 5% CO2, and that 5% O2 is superior to either 1% or 20% O2.

Animals↗

PubMed1995

Nuclear transfer of putative rabbit embryonic stem cells leads to normal blastocyst development.

Rabbit embryonic stem-like cells, characterized by embryoid body formation and differentiation into cell types representative of all three germ layers, were studied for their ability to promote early embryonic development after nuclear transfer. After culture of the reconstructed embryos, 23% (n = 35) developed successfully into morulae or blastocysts, compared with 34% (n = 62) for cloned embryos derived from nuclear transfer with embryonic blastomeres. The cloned embryos from the embryonic stem-like cells appeared normal, with an average of 26% inner cell mass cells, similar to that of control non-manipulated embryos (25%) or cloned embryos from blastomeres (25%). Thus, nuclear transfer of rabbit embryonic stem-like cells leads to early embryonic development that is indistinguishable from blastomere fusion. These results have implications for the development of gene targeting in a species (rabbit) that may be a more suitable model for studying certain human diseases. In addition, this technique may be applicable to other species from which putative embryonic stem cells have been derived, particularly agriculturally important animals.

Animals↗

PubMed1995

Repeatability of measurements on human, rabbit, and bull sperm by computer-assisted sperm analysis when comparing individual fields and means of 12 fields.

OBJECTIVE: To compare repeatability of measurements of human, rabbit, and bull sperm on two Hamilton Thorne Integrated Visual Optical System (IVOS) units, software version 10. DESIGN: Semen samples from seven normal human subjects, six rabbits, and eight bulls were obtained at regular intervals. The samples were diluted, two chambers were filled, and 12 fields were recorded, using high resolution recorders. Computer-assisted sperm analysis (CASA) was performed nearly simultaneously with two Hamilton Thorne IVOS units. SETTING: Reproduction Research Laboratories, Cornell University and Hamilton-Thorne Research, Beverly, Massachusetts. RESULTS: Optimal settings were established for evaluating by CASA sperm from three species. Fifteen variables were analyzed. The correlation coefficients for most variables characterizing sperm motion and concentration, when means of 12 fields were calculated, were 0.95 to 1.00. There were too few hyperactive sperm to obtain a reliable correlation for human sperm (r = 0.63) and repeatability of elongation was lower only for human sperm (r = 0.75). CONCLUSIONS: Two units of Hamilton Thorne IVOS, software version 10, were capable of providing nearly identical estimates of many CASA variables of human, rabbit, and bull sperm. Correlations for the paired estimates of many motion characteristics and sperm concentration usually exceeded 0.97.

Animals↗

PubMed1994

Surface alterations of the bovine oocyte and its investments during and after maturation and fertilization in vitro.

Surface characteristics of the bovine oocyte and its investments before, during, and after maturation, and fertilization in vitro were evaluated by scanning electron microscopy (SEM). Oocyte diameters were also measured during SEM analysis of the oocyte. The cumulus cells manifested a compact structure with minimal intercellular spaces among them in the immature oocytes. These became fully expanded with increased intercellular spaces after maturation in vitro, but contracted again after fertilization. The zona pellucida (ZP) showed a fibrous, open mesh-like structure in the maturing and matured oocytes. The size and number of meshes on the ZP decreased dramatically after fertilization. The vitelline surface of immature oocytes was characterized by distribution of tongue-shaped protrusions (TSPs) varying in density. After 10 and 22 hr of maturation incubation, oocyte surface microvilli (MV) increased to become the predominant surface structure, and TSPs decreased substantially. The vitelline surface of fertilized oocytes (at 6 and 20 hr) was similar to that of the matured oocytes, but unfertilized oocytes had less dense MV than did fertilized oocytes (at 20 hr). The diameter of the oocytes decreased from 99 to 80 microns during maturation and increased to 106 microns after insemination (P < 0.05). Membrane maturation was characterized by surface changes from a TSP-predominant pattern to a MV-predominant pattern. Thus, the bovine oocyte maturation process was found to involve the expansion of cumulus cells and the maturation of the ZP, which changes dramatically upon fertilization. Also, volumetric changes occurred in ooplasm processed for SEM following oocyte maturation and insemination.

Animals↗

PubMed1994

Synergistic effect of ethanol and cycloheximide on activation of freshly matured bovine oocytes.

Bovine follicular oocytes were collected from ovarian antral follicles (2 to 7 mm in diameter) from slaughtered cattle. They were matured in vitro (IVM) for 23 to 24 h and then activated. In Experiment 1, 4 concentrations of ethanol were compared. The activation rates of oocytes were 4, 12, 36 and 27%, respectively, following exposure for 7 min to 0, 5, 7 and 10% ethanol. In Experiment 2, 7% ethanol was tested with exposure times of 0, 5, 7.5 and 10 min, and 6, 32, 27 and 33% of the oocytes were activated, respectively. In Experiment 3 the synergistic effect of ethanol and electric pulse was compared within 4 treatments: A) 7% ethanol alone, B) electric pulse alone, C) ethanol first and then electric pulse treatment, and D) electric pulse first followed by ethanol exposure. Of the oocytes activated, 37, 31, 28 and 51%, respectively, were from Treatments A through D. In Experiments 4 and 5 the possible synergistic effect of ethanol and a protein synthesis inhibitor, cycloheximide, was studied within 4 treatments: A) parthenogenetic control with no activation treatment, B) ethanol alone, C) cycloheximide alone, and D) ethanol treatment followed by cycloheximide. The oocyte activation rates in Experiment 4 in Treatments A through D, respectively, were 9, 44, 43 and 84%. Corresponding values for development of oocytes to the 2 to 8-cell stage after culture for 3 d (Experiment 5) were 9, 20, 14 and 45%, respectively (P<0.05). In conclusion, exposure to 7% ethanol for 5 min followed by incubation with cycloheximide was the best activation treatment for bovine IVM oocytes.

Journal Article↗

PubMed1994

Relationships between administration of GnRH, body condition score and fertility in Holstein dairy cattle.

The objective of this study was to determine the effect of GnRH treatment in dairy cows differing in body condition score. A total of 2437 primiparous and multiparous cows was allocated randomly to receive either GnRH (100 microg) or placebo at the post partum exam (25 to 35 d) and at the first breeding. Complete records were available from 1906 cows: Group 1 (n = 535) placebo and placebo; Group 2 (n = 489) GnRH and placebo; Group 3 (n = 438) placebo and GnRH; and Group 4 (n = 444) GnRH and GnRH. The change of body condition score during early lactation was determined on a 1 to 5 scale (1 = emaciated to 5 = obese). Cows with a high body condition score (> or =3.0) at the first breeding had 8.8 fewer days to first service and 6.4 fewer days open but more services per conception (0.16) than cows with low body condition score (<3.0). Conception rate improved when GnRH was administered at the first breeding to cows with a body condition score of <3.0 at the first breeding regardless of parity. The administration of GnRH at the post partum exam decreased the conception rate in first lactation cows but was beneficial for cows in second and greater lactations. Thus the efficacy of GnRH may not be consistent in all parities and body condition groups.

Journal Article↗

PubMed1994

Analysis of polarity of bovine and rabbit embryos by scanning electron microscopy.

Cellular polarization during preimplantation development of the embryo is believed to be a crucial event in the transition of a zygote to a blastocyst stage embryo with morphologically and functionally differentiated cell types. While extensive studies have been done on polarity development in mouse embryos, little information is available in other species, particularly in cattle. The objective of this study was to examine the initiation of polarity by microvilli distribution on blastomeres of cattle and rabbit embryos using scanning electron microscopy (SEM). Bovine embryos were obtained by in vitro fertilization of in vitro-matured follicular oocytes. Rabbit embryos of various stages were collected from superovulated rabbits. Intact embryos and isolated blastomeres were examined in both species. Blastomeres from 1- to 8-cell embryos in both cattle and rabbits showed no polarity. The onset of transitional polarization of microvillous distribution occurred in some blastomeres of cattle embryos at the 9- to 15-cell stage; but typical, distinct polarity was not manifested until after the 16-cell stage with approximately 40% polar cells per embryo. In the rabbit blastomere polarity occurred one cell cycle later, with 46% polar cells per embryo after the 32-cell stage. The difference in cell numbers at the time polarity is evident is probably related to the different cell stages for embryo compaction and blastocyst formation in the two species.

Animals↗

PubMed1994

Semen quality and behavior of Holstein bulls exposed to estradiol-treated bulls for mounts.

The objectives were to test the effects of estradiol treatment of teaser bull mounts on sexual behavior and on quality and quantity of sperm obtained from sires managed as in large commercial AI breeding organizations. In a change-over design, the same teaser bulls were either untreated or treated with estradiol. Five semen-producing bulls were ejaculated twice per day on Tuesdays and Fridays after epididymal reserves were partially depleted. A 15-min period of continuous sexual preparation with three false mounts allowed was standard before each semen collection. All bulls were attracted to and licked the preputial area of the teaser bulls followed by the Flehmen response during the period of sexual preparation. Bulls usually completed the false mounts in < or = 15 min, and all thrusted vigorously with both hind feet moving forward synchronously at this time on 100% of the 80 semen collections. Major differences among bulls and between first and second ejaculates occurred in semen volume, semen concentration, and total sperm collected. An increase of 10% in total sperm output when bulls were exposed to treated teaser bulls could be of commercial benefit. The correlation between total time to first mount for the two ejaculates per bull each day and total sperm collected per bull per day was -.44. Thus, the shorter time to first mount may be useful as a low level predictor of higher sperm out-put per bull.

Animals↗

PubMed1993

Bovine oocyte development following different oocyte maturation and sperm capacitation procedures.

Various procedures have been reported for successful in vitro maturation and in vitro fertilization (IVM/IVF) of bovine follicular oocytes. Direct comparisons of these different recommended procedures have been rare. In this research, involving a total of 5,128 oocytes, a series of experiments were conducted to compare oocyte maturation, fertilization, and development in vitro with 2 maturation systems (with or without added hormones) and 3 types of sperm treatment procedures. Oocytes were collected from ovarian antral follicles (2-7 mm in diameter) within 3 hr after slaughter of cows or heifers. Those with intact or at least 4 layers of cumulus cells were selected for IVM/IVF. Oocytes were incubated for 22 hr in either Medium 199 with 7.5% fetal calf serum (M199 + FCS) alone or M199 + FCS with added hormones (M199 + FCS + H; oFSH 0.5 micrograms/ml, oLH 5.0 micrograms/ml, and E2 1.0 micrograms/ml) at 39 degrees C in 5% CO2 and 95% air. For IVF, frozen-thawed sperm were treated with either 0.1 microM calcium ionophore A23187 (A23187) for 1 min, or 10 or 100 micrograms/ml heparin (H10 or H100) for 15 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Nuclear transfer in cattle: effect of nuclear donor cells, cytoplast age, co-culture, and embryo transfer.

There are many factors affecting the efficiency of nuclear transfer technology. Some are evaluated here using our novel approach by enucleating oocytes at 20-22 hr after in vitro maturation (IVM), culturing the enucleated oocytes (cytoplasts) for 8-10 hr or 18-20 hr to gain activation competence and then conducting nuclear transfer. In the first experiment, we demonstrated that cumulus cell (CC) monolayer can support some cloned embryos to develop into morulae or blastocysts. Co-culture with CC and bovine oviduct epithelial cell (BOEC) monolayers resulted in no differences (P > 0.05) in supporting the development of cloned embryos (Experiment 2). When in vitro matured oocytes were enucleated at 22 hr after IVM followed by nuclear transfer 18-20 hr later, cleavage and morula or blastocyst development of the cloned embryos were similar to those resulting from the enucleated oocytes which had been matured in vivo (Experiment 3). Frozen embryos as nuclear donor cells worked equally well as fresh embryos for cloning in embryo development which was superior to IVF embryos (Experiment 4). However, fresh embryos resulted in a higher proportion (P < 0.05) of blastomere recovery than did frozen of IVF embryos. Finally, embryo transfer of cloned embryos from our procedure produced a viable calf, demonstrating the commercial value of this novel approach of the technology.

Animals↗

PubMed1993

Pluripotency of cultured rabbit inner cell mass cells detected by isozyme analysis and eye pigmentation of fetuses following injection into blastocysts or morulae.

Pluripotency of isolated rabbit inner cell masses (ICMs) and cultured (3 days) inner cell mass (ICM) cells was tested by injecting these donor cells into day 3.5 blastocysts (experiment 1) or day 3 morulae (experiment 2) to produce chimeric embryos. Injected (n = 107) and noninjected (n = 103) embryos were transferred to the opposite uterine horns of the same recipient females. Chimerism was determined by adenosine deaminase (ADA) isozyme analysis on fetal tissue and by eye pigmentation at midgestation. In experiment 1, 53% and 64%, respectively, of blastocysts injected with ICMs or cultured ICM cells developed to midgestation, compared with 52% and 48% for controls. Of these fetuses, four (31%) and one (6%), respectively, had ADA chimerism. In experiment 2, 38% and 62%, respectively, of the morulae injected with ICMs or cultured ICM cells developed to midgestation, compared with 46% and 56% for control morulae. Six (43%) chimeric fetuses from morulae injected with ICMs were detected by ADA analysis, but 12 (86%) chimeric fetuses were detected by eye pigmentation, indicating that eye pigmentation was a more sensitive marker for chimerism than our ADA assay. None of the 14 fetuses recovered after injecting morulae with cultured ICM cells were chimeric with either marker. No chimeras developed from control embryos. These studies demonstrate 1) that pregnancy rates are not compromised by injection of blastocysts or morulae with ICMs or cultured ICM cells, 2) that chimeric rabbit fetuses can be produced by injecting ICMs into either blastocysts or morulae, and 3) that cultured ICM cells can contribute to embryonic development when injected into blastocysts.

Animals↗

PubMed1993

Effect of sucrose, trehalose, hypotaurine, taurine, and blood serum on survival of frozen bull sperm.

Factorially arranged experiments were conducted to study the effects of adding sucrose and trehalose, known to have cryoprotective properties, blood serum, and the antioxidants taurine and hypotaurine on sperm motility after freezing and thawing at different rates. At sugar concentrations of 0.1 M, osmolality of the whole milk (WM) or egg yolk-Tris freezing medium (without glycerol) was about 370 mOsmol. Survival following freezing and thawing was reduced unless osmolality was corrected by adding pure water to reduce osmolality to about 280 mOsmol. Then the post-thaw percentages of motile sperm for control WM, and WM with 0.05 or 0.10 M sucrose, or 0.05 or 0.10 M trehalose, respectively, were 62, 55, 61, 57, and 62%. Thawing semen at 5 degrees C, versus 25 degrees C resulted in 64 versus 70% motile sperm (P < 0.05). Post-thaw survival of sperm stored at 25 degrees C for 24 h in trehalose-treated WM was superior to that in WM (P < 0.05). Hypotaurine and taurine had little effect on sperm survival. Up to 10% (v/v) of heated blood serum was generally beneficial, but gave more variable responses with different bulls. Sperm survival after cooling at -25 degrees C/min was slightly superior to that after cooling at -15 degrees C/min to -100 degrees C. The effects of the compounds studied on motility of frozen-thawed sperm were small, but if they protect sperm cell membranes, as reported for other types of membranes, they may assist sperm in surviving in the reproductive tract of the cow prior to fertilization.

Animals↗

PubMed1993

Effect of growth factors on morula and blastocyst development of in vitro matured and in vitro fertilized bovine oocytes.

Growth factors were studied as a means of increasing the development of in vitro matured (IVM) and in vitro fertilized (IVF) oocytes into morulae or blastocysts. Cell numbers of blastocysts were also counted. In Experiment 1, 2- to 8-cell embryos derived from bovine IVM/IVF oocytes were randomly allotted to one of 3 culture groups: a) synthetic oviduct fluid (SOF); b) SOF + 10 ng/ml epidermal growth factor (EGF); or c) SOF + 100 ng/ml EGF; all 3 culture media contained 10% fetal bovine serum. Culture resulted in 12%, 23% and 14% (P>0.05), respectively, developing into morulae and blastocysts. In Experiment 2, 5 ng/ml of transforming growth factor B (1) (TGF B (1)) added to CR(1aa) medium containing BSA increased the percentage of blastocysts to 56% vs 40% for the control (P<0.05). In Experiment 3, EGF and TGF B(1), added singly and in combination to CR(1aa) did not produce a synergistic effect. More embryos developed into morulae and blastocysts (45%) in a bovine oviduct epithelial co-culture than in any other treatment except in CR(1aa) + EGF (34%; P>0.05). In Experiment 4, 0, 1 and 5 ng/ml of platelet derived growth factor (PDGF) added to CR(1aa) yielded 39%, 70% and 52% morulae and blastocysts, respectively (P<0.05). Cell number was not increased, indicating that growth factors can increase the proportion of embryos that develop into morulae and blastocysts without an increase in the cell number.

Journal Article↗