Secretion profiles from in vitro cultured follicles, isolated from fresh prepubertal and adult mouse ovaries or frozen–thawed prepubertal mouse ovaries

Zygote ◽  
2011 ◽  
Vol 20 (2) ◽  
pp. 181-192 ◽  
Author(s):  
B. Dorphin ◽  
M. Prades-Borio ◽  
A. Anastacio ◽  
P. Rojat ◽  
C. Coussieu ◽  
...  
Keyword(s):  
Adult Mouse ◽  
Ovarian Tissue ◽  
New Technology ◽  
Growth Curves ◽  
Oocyte Quality ◽  
Inhibin B ◽  
Follicular Growth ◽  
Hormone Production

SummaryIn vitro folliculogenesis could be a new technology to produce mature oocytes from immature follicles that have been isolated from cryopreserved or fresh ovarian tissue. This technique could also be a tool for evaluation of oocyte quality and/or for determination of follicular parameters during follicular growth. Our objective was to characterize in mice the secretion profiles of follicles that had been isolated mechanically during in vitro follicular growth and in relation to the growth curve. Early preantral follicles from fresh prepubertal and adult mouse ovaries or frozen–thawed prepubertal mouse ovaries were cultured individually in microdrops under oil for 12 days. Each day, two perpendicular diameters of the follicles were measured. From day-3 to day-12 of culture, culture medium was collected and preserved for determination of inhibin B, anti-Müllerian hormone (AMH) and estradiol levels. At the end of the culture, after maturation, the status of the oocyte was evaluated. Follicular growth and their individual hormone production did not always correlate. Inhibin B was never secreted from follicles of less than 200 μm diameter, whether the follicles were examined when fresh or after freezing–thawing. Estradiol secretion was never observed in frozen–thawed follicles. AMH was mainly secreted between day-3 and day-9. Despite similar morphological aspects at the start of culture, follicles selected for in vitro folliculogenesis were found to be heterogeneous and differed in their ability to grow and to produce hormones, even if they had similar growth curves. Follicles from frozen–thawed ovaries developed slowly and produced fewer hormones than freshly collected follicles.

Factors affecting folliculogenesis in ruminants

1999 ◽  
Vol 68 (2) ◽  
pp. 257-284 ◽  
Author(s):  
R. Webb ◽  
R. G. Gosden ◽  
E. E. Telfer ◽  
R. M. Moor
Keyword(s):  
New Technologies ◽  
Ovarian Function ◽  
Major Gene ◽  
Follicular Development ◽  
Oocyte Quality ◽  
Ovulation Rate ◽  
Follicular Growth ◽  
Stages Of Development ◽  
Hormone Production ◽  
Factors Affecting

AbstractThis review addresses the reasons for the lack of progress in the control of superovulation and highlights the importance of understanding the mechanisms underlying follicular development. The present inability to provide large numbers of viable embryos from selected females still restricts genetic improvement, whilst variability in ovarian response to hormones limit the present capacity for increasing reproductive efficiency.Females are born with a large store of eggs which rapidly declines as puberty approaches. If these oocytes are normal then there is scope for increasing the reproductive potential of selected females. Oocytes must reach a certain size before they can complete all stages of development and the final changes that occur late in follicular development. It is likely that oocytes that do not produce specific factors at precise stages of development will not be viable. Hence, it is important to characterize oocyte secreted factors since there are potential indicators of oocyte quality.The mechanisms that determine ovulation rate have still not been fully elucidated. Indeed follicular atresia, the process whereby follicles regress, is still not known. A better understanding of these processes should prove pivotal for the synchronization of follicular growth, for more precise oestrous synchronization and improved superovulatory response.Nutrition can influence a whole range of reproductive parameters however, the pathways through which nutrition acts have not been fully elucidated. Metabolic hormones, particularly insulin and IGFs, appear to interact with gonadotrophins at the level of the gonads. Certainly gonadotropins provide the primary drive for the growth of follicles in the later stages of development and both insulin and IGF-1, possibly IGF-2, synergize with gonadotrophins to stimulate cell proliferation and hormone production. More research is required to determine the effects of other growth factors and their interaction with gonadotropins.There is evidence, particularly from studies with rodents, that steroids can also modulate follicular growth and development, although information is very limited for ruminants. There may be a rôle for oestrogens in synchronizing follicular waves, to aid in oestrous synchronization regimes and for removing the dominant follicle to achieve improved superovulatory responses. However more information is required to determine whether these are feasible approaches.Heritability for litter size is higher in sheep than in cattle. Exogenous gonadotropins are a commercially ineffective means of inducing twinning in sheep and cattle. Although there are differences in circulating gonadotropin concentrations, the mechanism(s) responsible for the high ovulation appear to reside essentially within the ovaries. The locus of the Booroola gene, a major gene for ovulation rate, has been established but not specifically identified. However sheep possessing major genes do provide extremely valuable models for investigating the mechanisms controlling ovulation rate, including a direct contrast to mono-ovulatory species such as cattle.In conclusion, the relationship between oocyte quality, in both healthy follicles and those follicles destined for atresia, must be resolved before the future potential for increasing embryo yield can be predicted. In addition, a greater understanding of the factors affecting folliculogenesis in ruminants should ensure that the full benefits ensuing from the precise control of ovarian function are achieved. The improved use of artificial insemination and embryo transfer that would ensue from a greater understanding of the processes of folliculo genesis, coupled with the new technologies of genome and linkage mapping, should ensure a more rapid rate of genetic gain.

scholarly journals The Calgary Biofilm Device: New Technology for Rapid Determination of Antibiotic Susceptibilities of Bacterial Biofilms

1999 ◽  
Vol 37 (6) ◽  
pp. 1771-1776 ◽  
Author(s):  
H. Ceri ◽  
M. E. Olson ◽  
C. Stremick ◽  
R. R. Read ◽  
D. Morck ◽  
...  
Keyword(s):  
Antibiotic Susceptibility ◽  
Clinical Laboratory ◽  
Rapid Determination ◽  
New Technology ◽  
Growth Curves ◽  
National Committee ◽  
Standard Group ◽  
Antibiotic Susceptibilities ◽  
Significant Difference

Determination of the MIC, based on the activities of antibiotics against planktonic bacteria, is the standard assay for antibiotic susceptibility testing. Adherent bacterial populations (biofilms) present with an innate lack of antibiotic susceptibility not seen in the same bacteria grown as planktonic populations. The Calgary Biofilm Device (CBD) is described as a new technology for the rapid and reproducible assay of biofilm susceptibilities to antibiotics. The CBD produces 96 equivalent biofilms for the assay of antibiotic susceptibilities by the standard 96-well technology. Biofilm formation was followed by quantitative microbiology and scanning electron microscopy. Susceptibility to a standard group of antibiotics was determined for National Committee for Clinical Laboratory Standards (NCCLS) reference strains: Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, andStaphylococcus aureus ATCC 29213. Growth curves demonstrated that biofilms of a predetermined size could be formed on the CBD at specific time points and, furthermore, that no significant difference (P > 0.1) was seen between biofilms formed on each of the 96 pegs. The antibiotic susceptibilities for planktonic populations obtained by the NCCLS method or from the CBD were similar. Minimal biofilm eradication concentrations, derived by using the CBD, demonstrated that for biofilms of the same organisms, 100 to 1,000 times the concentration of a certain antibiotic were often required for the antibiotic to be effective, while other antibiotics were found to be effective at the MICs. The CBD offers a new technology for the rational selection of antibiotics effective against microbial biofilms and for the screening of new effective antibiotic compounds.

Live Birth After an Autologous Platelet-Rich Plasma Ovarian In Vitro Activation and Bone Marrow Stem Cells Transplantation in a Premature Ovarian Failure Case Report

2021 ◽  
Author(s):  
Aleksandar Ljubić ◽  
Tatjana Božanović ◽  
Andrea Pirkovic-Cabarkapa ◽  
Andjela Perovic ◽  
Dušica Ljubić ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Ovarian Function ◽  
Platelet Rich Plasma ◽  
Ovarian Tissue ◽  
Follicular Growth ◽  
Hormonal Function ◽  
Autologous Platelet Rich Plasma ◽  
Natural Conception ◽  
Autologous Platelet

Abstract Background: Patients with premature ovarian failure (POF) exhibit a diminished ovarian reserve and hormonal dysfunction. Case presentation: We aimed to restore normal hormonal function and folliculogenesis in a 31-year-old patient with POF, who had been amenorrheic for two years. We designed and performed three different ovarian regeneration procedures for three consecutive years, from 2015 to 2017: 1) intraovarian injection of activated autologous platelet-rich plasma (PRP); 2) activated autologous PRP and bone marrow-derived stem cell injection into the ovaries (SEGO); 3) ovarian cortical tissue resection and fragmentation; and in vitro ovarian tissue activation with autologous PRP and bone marrow stem cell retransplantation into the ovaries (the SEGOVA). The patient exhibited no improvement following the PRP treatment. The patient regained regular menstrual cycles after the SEGO procedure, although no follicular growth was observed yet. One month after the SEGOVA procedure, follicular growth was detected, and the patient underwent several stimulation protocols without obtaining oocytes. Eight months after the SEGOVA, the patient underwent in vitro fertilization (IVF) in a spontaneous cycle, when an oocyte of good quality was retrieved. Following intracytoplasmic sperm injection (ICSI), the oocyte failed to be fertilized. Eleven months after the SEGOVA, the patient reported a spontaneous pregnancy via natural conception. Pregnancy resulted in birth at term by uncomplicated vaginal delivery. After three different ovarian rejuvenation procedures, normal hormonal function and follicular growth were restored in the patient with POF, and the patient had a successful natural pregnancy following the last SEGOVA procedure. Although no ovarian function was detected after the first two procedures, they may have contributed to the outcomes from the SEGOVA procedure, a treatment that showed promising results in recovering ovarian function in patients with POF. Conclusions: It cannot be ruled out that ovarian rejuvenation with bone marrow‑derived stem cells and autologous growth factors, together with ovarian tissue fragmentation, took time to exhibit its effects and contributed to the final result – a successful natural conception and pregnancy.

Human adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: Hormone production

2006 ◽  
Vol 217 (1) ◽  
pp. 114-124 ◽  
Author(s):  
Markus Hecker ◽  
John L. Newsted ◽  
Margaret B. Murphy ◽  
Eric B. Higley ◽  
Paul D. Jones ◽  
...  
Keyword(s):  
Hormone Production ◽  
H295r Cells

EFFECTS OF PITUITARY HORMONES ON PROGESTATIONAL HORMONE PRODUCTION BY THE RABBIT OVARY IN VIVO AND IN VITRO

1966 ◽  
Vol 35 (1) ◽  
pp. 53-63 ◽  
Author(s):  
J. H. DORRINGTON ◽  
R. KILPATRICK
Keyword(s):  
Ovarian Tissue ◽  
Venous Blood ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Hormone Production ◽  
Stimulatory Action ◽  
Steroid Synthesis ◽  
Ovine Prolactin

SUMMARY Ovine luteinizing hormone (LH) increased the output of progestational steroids (20α-hydroxypregn-4-en-3-one and progesterone) in rabbit ovarian venous blood. Similar increases were found with ovine follicle-stimulating hormone (FSH) and growth hormone, but much larger amounts were necessary. Ovine prolactin was without effect. The increased output was due to increased synthesis and not only to release of stored steroids. Synthesis of these progestational steroids was stimulated by LH incubated with rabbit ovarian tissue. The stimulation produced by FSH was probably due to contamination by LH since the log dose-response lines for LH and FSH were parallel, and FSH was approximately 100 times less active than LH. Ovine prolactin had no stimulatory activity in concentrations up to 20 μg./ml. The stimulatory action of LH was unrelated to the presence of corpora lutea. Separated corpora lutea showed only a slight response to LH, whereas the response of interstitial tissue was similar to that found with undissected ovaries. Hence LH caused progestational steroid synthesis by stimulating the ovarian interstitial tissue.

Influence of follicle-stimulating hormone concentrations on the integrity and development of bovine follicles cultured in vitro

Zygote ◽  
2018 ◽  
Vol 26 (5) ◽  
pp. 417-423 ◽  
Author(s):  
C. Bizarro-Silva ◽  
M.M. Santos ◽  
J.R. Gerez ◽  
S.M. González ◽  
L.A. Lisboa ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Follicle Stimulating Hormone ◽  
Follicular Development ◽  
Follicular Growth ◽  
Minimal Essential Medium ◽  
Preantral Follicles ◽  
Mean Diameter ◽  
Stimulating Hormone

SummaryThis study investigated the in vitro culture of bovine follicles included in ovarian tissue for 2 or 6 days (D2 or D6), with the addition of different concentrations of follicle-stimulating hormone (FSH) (0, 10, 50, 100 or 200 ng/ml). Data were compared for follicular development, morphological integrity and diameter of follicles and oocytes. Ovaries (n = 10) from Nelore cows (n = 5) were divided into fragments (n = 11 per ovary) and were immediately fixed in Bouin’s solution (D0) or were individually cultured for 2 or 6 days in one of the described concentrations of FSH and then processed for histology. Compared with the rates of follicular development at D2 for minimal essential medium (MEM) (75.0%) and 50 ng/ml of FSH (71.1%), the best rates of follicular development at D2 were obtained with 10 (84.7%), 100 (87.5%) and 200 ng/ml of FSH (85.0%; P<0.05). After 6 days of cultivation, there were no differences among treatments regarding follicular growth. The morphological integrity of preantral follicles was better maintained by 100 ng/ml FSH for 2 and 6 days of cultivation (51.2 and 40.4%, respectively; P<0.05) than that for MEM (D2: 30.9%, D6: 20.8%), 10 (D2: 39.2%, D6: 22.8%), 50 (D2: 30.4%, D6: 28.8%) and 200 ng/ml FSH (D2: 45.2%, D6: 36.8%). FSH at 100 ng/ml provided the highest mean diameter averages: 34.5±10.8 µm at D2 and 33.2±12.5 µm at D6 (P<0.05). We concluded that the medium supplemented with 100 ng/ml FSH during in vitro culture provided appropriate conditions for the development and morphological integrity of preantral follicles in cattle.

In vivo and in vitro strategies to support caprine preantral follicle development after ovarian tissue vitrification

2018 ◽  
Vol 30 (8) ◽  
pp. 1055 ◽  
Author(s):  
N. J. Donfack ◽  
K. A. Alves ◽  
B. G. Alves ◽  
R. M. P. Rocha ◽  
J. B. Bruno ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Preantral Follicle ◽  
Hormone Production ◽  
Treatment Groups ◽  
Morphology Development ◽  
Fresh Culture

The aim of the present study was to compare fresh and vitrified goat ovarian tissue after autotransplantation and in vitro culture. Adult goats were completely ovariectomised and each ovarian pair was sliced and distributed among six different treatment groups: fresh control, fresh transplant, fresh culture, vitrified control, vitrified transplant and vitrified culture. Follicular morphology, development, growth, density, revascularisation and hormone production were evaluated in all groups. Three antral follicles (two in the fresh transplant and one in the vitrified transplant groups) were observed on the surface of the graft 90 days after transplantation. The percentage of morphologically normal follicles was similar in the fresh control, fresh transplant and vitrified transplant groups. The percentage of developing (transition, primary and secondary) follicles was higher after in vitro culture of fresh or vitrified tissue. Transplantation resulted in a lower follicle density. Serum oestradiol concentrations remained constant during the entire transplantation period. In contrast, progesterone production decreased significantly. Expression of CD31 mRNA was lower in fresh culture. In conclusion, restoration of goat ovarian function can be successfully achieved following transplantation of both fresh and vitrified goat ovarian tissue. However, transplantation induced higher follicle loss than in vitro culture.

Aloe vera increases mRNA expression of antioxidant enzymes in cryopreserved bovine ovarian tissue and promotes follicular growth and survival after in vitro culture

Cryobiology ◽  
2021 ◽  
Author(s):  
Francisco das Chagas Costa ◽  
Erlândia Márcia Vasconcelos ◽  
Venância Antônia Nunes Azevedo ◽  
Laís Raiane Feitosa Melo Paulino ◽  
Mônica Dias Soares ◽  
...  
Keyword(s):  
Antioxidant Enzymes ◽  
In Vitro Culture ◽  
Mrna Expression ◽  
Ovarian Tissue ◽  
Aloe Vera ◽  
Follicular Growth ◽  
Growth And Survival

scholarly journals An Attenuating Mutation in nsP1 of the Sindbis-Group Virus S.A.AR86 Accelerates Nonstructural Protein Processing and Up-Regulates Viral 26S RNA Synthesis

2003 ◽  
Vol 77 (2) ◽  
pp. 1149-1156 ◽  
Author(s):  
Mark T. Heise ◽  
Laura J. White ◽  
Dennis A. Simpson ◽  
Christopher Leonard ◽  
Kristen A. Bernard ◽  
...  
Keyword(s):  
Sindbis Virus ◽  
Rna Synthesis ◽  
Adult Mouse ◽  
Nonstructural Protein ◽  
Growth Curves ◽  
Protein Processing ◽  
Wild Type ◽  
Rnase Protection ◽  
Nonstructural Protein 1

ABSTRACT The Sindbis-group alphavirus S.A.AR86 encodes a threonine at nonstructural protein 1 (nsP1) 538 that is associated with neurovirulence in adult mice. Mutation of the nsP1 538 Thr to the consensus Ile found in nonneurovirulent Sindbis-group alphaviruses attenuates S.A.AR86 for adult mouse neurovirulence, while introduction of Thr at position 538 in a nonneurovirulent Sindbis virus background confers increased neurovirulence (M. T. Heise et al., J. Virol. 74:4207-4213, 2000). Since changes in the viral nonstructural region are likely to affect viral replication, studies were performed to evaluate the effect of Thr or Ile at nsP1 538 on viral growth, nonstructural protein processing, and RNA synthesis. Multistep growth curves in Neuro2A and BHK-21 cells revealed that the attenuated s51 (nsP1 538 Ile) virus had a slight, but reproducible growth advantage over the wild-type s55 (nsP1 538 Thr) virus. nsP1 538 lies within the cleavage recognition domain between nsP1 and nsP2, and the presence of the attenuating Ile at nsP1 538 accelerated the processing of S.A.AR86 nonstructural proteins both in vitro and in infected cells. Since nonstructural protein processing is known to regulate alphavirus RNA synthesis, experiments were performed to evaluate the effect of Ile or Thr at nsP1 538 on viral RNA synthesis. A combination of S.A.AR86-derived reporter assays and RNase protection assays determined that the presence of Ile at nsP1 538 led to earlier expression from the viral 26S promoter without affecting viral minus- or plus-strand synthesis. These results suggest that slower nonstructural protein processing and delayed 26S RNA synthesis in wild-type S.A.AR86 infections may contribute to the adult mouse neurovirulence phenotype of S.A.AR86.

scholarly journals Description of the Follicular Fluid Cytokine and Hormone Profiles in Human Physiological Natural Cycles

2020 ◽  
Author(s):  
Marie-Pierre Piccinni ◽  
Rossella Vicenti ◽  
Federica Logiodice ◽  
Raffaella Fabbri ◽  
Ornela Kullolli ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Ovarian Tissue ◽  
Oocyte Quality ◽  
Cytokine Profile ◽  
Ovarian Tissue Cryopreservation ◽  
Regulation System ◽  
Follicle Development ◽  
Vitro Fertilization ◽  
Natural Cycles

Abstract Purpose Exogenous gonadotrophins administration during in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles could significantly alter the endogenous follicular regulation system and could influence oocyte quality. The analysis of the follicular fluid (FF) cytokine and hormone profiles in physiological natural cycles is crucial to appreciate the role of FF milieu on follicle development. So far, the FF cytokine profile has been analyzed only in controlled ovarian stimulation cycles and in modified natural cycles. Our study defines, in physiological natural cycles, the cytokine and hormone profiles of individual FF aspirated from antral follicles. Methods A total of 203 FFs obtained from 83 women with regular menstrual cycles undergoing ovarian tissue cryopreservation were analyzed: 115 FFs from Group 1 (10 to 29 years of age) and 88 FFs from Group 2 (30 to 40 years of age). In individual FF, 27 cytokines were measured with xMAP technology, and progesterone, estrone, estradiol, testosterone, androstenedione concentrations were determined by liquid chromatography–tandem mass spectrometry. Results FF hormone profiles were not different in follicular and luteal phase, suggesting that FF hormones are regulated independently of the endogenous gonadotrophins—possibly because 74% of the punctured follicles, which were ≤6 mm, did not require cyclic pituitary function. The follicle size was influenced not only by the FF cytokine profile but also by the FF hormone profile, both of which are dependent on age. Main Conclusions In physiological natural cycles, FF hormones seems to be regulated independently of the endogenous gonadotropins. Age influences FF hormone and cytokine profiles and the compelling relationship between FF hormones and FF cytokines could influence the follicle development.

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