Effect of ovariectomy and graft position on cryopreserved common wombat (Vombatus ursinus) ovarian tissue following xenografting to nude mice

2003 ◽  
Vol 15 (6) ◽  
pp. 333 ◽  
Author(s):  
M. Cleary ◽  
M. C. J. Paris ◽  
J. Shaw ◽  
G. Jenkin ◽  
A. Trounson
Keyword(s):  
Ovarian Tissue ◽  
Polar Body ◽  
Antral Follicle ◽  
Follicle Development ◽  
Hormonal Stimulation ◽  
Kidney Capsule ◽  
Common Wombat ◽  
The One ◽  
Endangered Animals

Ovarian tissue xenografting may be applied to increase the population size of rare or endangered animals. However, optimal grafting conditions, such as graft position and recipient hormonal status, are yet to be established. The present study, using common wombat ovarian tissue, showed that development of xenografted ovarian tissue to the antral follicle stage can be achieved irrespective of graft position. However, increased graft recovery rates and follicle survival were evident after grafting under the kidney capsule compared with grafting to subcutaneous sites. No increase in follicle development was observed after placing grafts both under the kidney capsule and subcutaneously in the one recipient compared with grafts placed under the kidney capsule alone or subcutaneously alone. Removal of the recipient’s own ovaries at the time of grafting accelerated graft follicle development, with antral follicles seen by Week 12 after grafting compared with by Week 16 in recipients that retained their own ovaries. More oocytes were collected from xenograft recipients receiving hormonal stimulation before collection compared with non-stimulated recipients. No oocytes were mature (extruded a polar body) at the time of collection or after a subsequent period of in vitro maturation. This is the first study to demonstrate that antral follicle development can occur and oocytes can be collected from xenografted common wombat ovarian tissue.

Influence of hormone environment and donor age on cryopreserved common wombat (Vombatus ursinus) ovarian tissue xenografted into nude mice

2004 ◽  
Vol 16 (7) ◽  
pp. 699 ◽  
Author(s):  
M. Cleary ◽  
J. M. Shaw ◽  
G. Jenkin ◽  
A. O. Trounson
Keyword(s):  
Nude Mice ◽  
Ovarian Tissue ◽  
Follicle Development ◽  
Kidney Capsule ◽  
Oocyte Collection ◽  
Common Wombat ◽  
Tissue Grafts ◽  
Gonadal Status

Developmentally competent oocytes can be collected from xenografted ovarian tissues; however, optimal xenograft conditions need to be established for this technique to be of use in assisted reproduction. In the present study, common wombat ovarian tissue was xenografted under the kidney capsule of nude mice to clarify the role of recipient gonadal status and donor tissue age on graft establishment, follicle development and oocyte recovery. Eighty-nine per cent of all grafts were recovered; of these, 78% contained growing follicles. In female graft recipients, follicle development to the antral stage occurred earlier in ovariectomised recipients compared with intact graft recipients. Similarly, follicle development occurred earlier in recipients of pouch young ovarian tissue grafts when compared with subadult xenografts. Follicle development proceeded to the antral stage in subadult grafts placed under the kidney capsule of male recipient mice, albeit at a slower rate than subadult grafts placed in female recipients. Oocytes were collected from grafts placed in female and male recipients, but no mature oocytes were observed at the time of collection, nor could these oocytes be matured in vitro. The present study demonstrated that common wombat pouch young tissue xenografted to female recipient mice, and subadult ovarian tissue xenografted to male recipient mice, can develop to the antral stage and can therefore facilitate oocyte collection. However, mature oocytes were not obtained using the current protocol.

328 IN VITRO MATURATION OF OOCYTES FROM Canindé GOATS SUBMITTED TO DIFFERENT HORMONAL TREATMENTS FOR OVARIAN STIMULATION

2010 ◽  
Vol 22 (1) ◽  
pp. 320
Author(s):  
K. C. Almeida ◽  
A. F. Pereira ◽  
A. S. Alcântara Neto ◽  
S. R. G. Avelar ◽  
F. C. Sousa ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Polar Body ◽  
Hormonal Treatment ◽  
Oocyte Quality ◽  
Exact Test ◽  
Gentamicin Sulfate ◽  
First Polar Body ◽  
The One ◽  
Hormonal Treatments

Oocyte IVM is a long process during which oocytes acquire their ability to support the stages of development in a stepwise manner, ultimately reaching activation of the embryonic genome. The overall success of this process can be affected by factors such as hormonal treatment for ovarian stimulation. Thus, the current study aims to evaluate the possible effects of the ovarian stimulatory protocols on the goat oocyte quality and IVM rate. Adult and cyclic Canindé goats were heat-synchronized by means of intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) inserted for 11 days coupled with a luteolytic injection of 50 μg cloprostenol (Ciosin, Coopers, São Paulo, Brazil) in the 8th day of treatment. The ovarian stimulation was carried out using one of the following protocols: a) standard multi-doses (MD) with 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five injections (30/30; 20/20; 20 mg) at 12 h intervals (n = 18); b) three- doses (TD) with 120 mg pFSH administered in three injections (60; 40; 20 mg) at 24 h intervals (n = 17); c) one shot (OD) of 70 mg pFSH plus 200 IU of eCG (Novormon, Syntex) administered 36 h before sponge removal (n = 17). In MD andTD groups, the pFSH injections started in Day 8 of progestagen treatment. The follicles were aspirated just after the sponge removal using laparoscopic oocyte recovery (LOR). This procedure was performed with a 22-gauge needle and a vacuum pump at 30 mmHg. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin sulfate (40 μg mL-1). COCs were classified as grade I, II, III, or IV based on visual criteria (Baldassarre H et al. 2003 Theriogenology 56, 831-839). Good quality oocytes (grade I and II) were incubated in TCM-199 supplemented with cysteamine (100 μM), EGF (10 ng mL-1) and gentamicin sulfate (40 μgm L-1) at 38.5°C in a humidified atmosphere with 5% CO2 in air for 24 h. Oocyte maturation was assessed by the visualization of first polar body under inverted microscope. Data were expressed as percentages and analyzed using the Fischer’s exact test. No statistical differences among hormonal treatments (P > 0.05) were observed for the percentage of the good quality oocytes, with 70.4 ± 3.0% of COCs graded in I and II. The IVM rate inTD (31.4%) was statistically lower than MD (31.4% v. 46.5%, P = 0.04) group. However, no significant differences (P = 0.89) were observed between OD (45.2%) and MD groups. Thus, current results indicate that oocyte production for IVM can be facilitated using ovarian stimulation with the one shot FSH/eCG regime without affecting meiotic competence. In summary, OD and MD treatments can be used for oocyte IVM in an embryo production programme in Canindé goats. This study was supported by the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

scholarly journals The activin-follistatin system and in vitro early follicle development in goats

2006 ◽  
Vol 189 (1) ◽  
pp. 113-125 ◽  
Author(s):  
J R V Silva ◽  
T Tharasanit ◽  
M A M Taverne ◽  
G C van der Weijden ◽  
R R Santos ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Primordial Follicle ◽  
Activin A ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Follicle Development ◽  
Cortical Tissue ◽  
Primary Follicle ◽  
Growth And Survival ◽  
Primordial Follicles

The aim of the present study was to investigate the effects of activin-A and follistatin on in vitro primordial and primary follicle development in goats. To study primordial follicle development (experiment 1), pieces of ovarian cortex were cultured in vitro for 5 days in minimal essential medium (MEM) supplemented with activin-A (0, 10 or 100 ng/ml), follistatin (0, 10 or 100 ng/ml) or combinations of the two. After culture, the numbers of primordial follicles and more advanced follicle stages were calculated and compared with those in non-cultured tissue. Protein and mRNA expression of activin-A, follistatin, Kit ligand (KL), growth differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15) in non-cultured and cultured follicles were studied by immunohistochemistry and PCR. To evaluate primary follicle growth (experiment 2), freshly isolated follicles were cultured for 6 days in MEM plus 100 ng/ml activin-A, 100 ng/ml follistatin or 100 ng/ml activin-A plus 200 ng/ml follistatin. Morphology, follicle and oocyte diameters in cultured tissue and isolated follicles before and after culture were assessed. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) reactions were performed to study DNA fragmentation in follicles. In experiment 1, it was found that goat primordial follicles were activated to develop into more advanced stages, i.e. intermediate and primary follicles, during in vitro culture, but neither activin-A nor follistatin affected the number of primordial follicles that entered the growth phase. Activin-A treatment enhanced the number of morphologically normal follicles and stimulated their growth during cortical tissue culture. The effects were, however, not counteracted by follistatin. The follicles in cultured goat tissue maintained their expression of proteins and mRNA for activin-A, follistatin, KL, GDF-9 and BMP-15. Fewer than 30% of the atretic follicles in cultured cortical tissue had TUNEL-positive (oocyte or granulosa) cells. Activin-A did not affect the occurrence of TUNEL-positive cells in follicles within cortical tissue. In experiment 2, addition of activin-A to cultured isolated primary follicles significantly stimulated their growth, the effect being counteracted by follistatin. Absence of such a neutralizing effect of follistatin in the cultures with ovarian cortical tissue can be due to lower dose of follistatin used and incomplete blockage of activin in these experiments. In contrast to cortical enclosed atretic follicles, all atretic follicles that had arisen in cultures with isolated primary follicles had TUNEL-positive cells, which points to differences between isolated and ovarian tissue-enclosed follicles with regard to the followed pathways leading to their degeneration. In summary, this in vitro study has demonstrated that cultured goat primordial follicles are activated to grow and develop into intermediate and primary follicles. During in vitro culture, the follicles maintain their ability to express activin-A, follistatin, KL, GDF-9 and BMP-15. The in vitro growth and survival of activated follicles enclosed in cortical tissue and the in vitro growth of isolated primary follicles are stimulated by activin-A.

scholarly journals Supplementation of c-type natriuretic peptide during the whole in vitro growth period benefits the development of murine secondary follicles

2020 ◽  
Author(s):  
Ang Li ◽  
Haixia Cao ◽  
Hongxia Li ◽  
Ruijiao Li ◽  
Huaixiu Wang ◽  
...  
Keyword(s):  
Natriuretic Peptide ◽  
Ovulation Induction ◽  
Germinal Vesicle ◽  
Antral Follicle ◽  
Control Group ◽  
Follicle Development ◽  
In Vitro Growth ◽  
Germinal Vesicle Breakdown ◽  
Preovulatory Follicles

Abstract Background Supplementation of c-type natriuretic peptide (CNP) in the culture medium shortly before in vitro maturation (IVM) has been reported to be effective in delaying meiotic resumption of murine oocyte. The present study investigated the effect of CNP supplementation during the whole period of in vitro growth (IVG) on the development of murine secondary ovarian follicles.Methods Late secondary ovarian follicles isolated from ovaries of Kunming mice were cultured in vitro with and without supplementation of CNP. In experiment 1, CNP was supplemented at the early stage and the follicle development was evaluated. In experiment 2 and 3, CNP was supplemented during the whole period of IVG. In experiment 2, follicle development and oocyte maturity were evaluated. In group 3, follicle development and rate of cleaved embryos after in vitro fertilization (IVF) was assessed.Results In control group in all 3 experiments, granulosa cells migrated from within follicle and adhered to the plate at different degrees. The follicles flattened and could not reach antral stage. About 39.8% (39/98) of the oocytes ovulated nakedly. As no antral follicle was obtained, IVF was not performed in control group in experiment 3. In experiment group in all 3 experiments, no migration of guanulosa cells was observed and the follicles grew three-dimensionally. Ovulation of naked oocyte decreased substantially. The rate of antral stage follicle were 45% (18/40) in experiment 1. This parameter was 75.9% (44/58) in experiment 2 and 3 combined. In experiment 2, in preovulatory follicles without ovulation induction, oocytes at germinal vesicle (GV) stage and germinal vesicle breakdown (GVBD) stage were 87.5% (14/16) and 12.5% (2/16), respectively. In preovulatory follicles with ovulation induction, no GV stage oocyte was retrieved, oocytes at GVBD and metaphase II (MII) stage were 50% (8/16), respectively. In experiment 3, among 18 follicles cultured, 12 cumulus-oocyte complexes (COC) ovulated automatically after ovulation induction. Eleven oocytes were fertilized and cleaved. Compared with control groups, the follicle development assessed by naked oocyte ovulation and follicle stage (preantral follicle and antral follicle) in experiment groups were significantly superior (p<0.0001). CNP effectively maintained oocytes’ meiotic arrest and enhanced fertilization competency.Conclusions The supplementation of CNP in culture system of murine late secondary follicle during the whole period of IVG could sustain the 3-dimensional structure of follicle, increase the antral formation rate. As a result, the oocyte’s competency to be fertilized was greatly improved.

Co-culture embedded in cumulus clumps promotes maturation of denuded oocytes and reconstructs gap junctions between oocytes and cumulus cells

Zygote ◽  
2012 ◽  
Vol 21 (3) ◽  
pp. 231-237 ◽  
Author(s):  
Guixue Feng ◽  
Deshun Shi ◽  
Shufang Yang ◽  
Xiaoli Wang
Keyword(s):  
Gap Junctions ◽  
Lucifer Yellow ◽  
Ovarian Tissue ◽  
Polar Body ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Control Group ◽  
First Polar Body ◽  
Cytoplasmic Maturation

SummaryThe present study was undertaken to establish an effective method for in vitro maturation (IVM) of denuded oocytes (DOs) by simulating the ovarian three-dimensional status in vivo using buffalo ovarian tissues or cumulus cells, so as to provide a model for investigating the mechanisms of oocyte maturation. Buffalo cumulus–oocyte complexes from ovaries taken at slaughter were denuded by pipetting, and then allocated randomly into four groups for IVM by direct culture in maturation medium (M1, control group), co-culture with a monolayer of cumulus cells (M2), embedded in cumulus cell clumps (M3) and ovarian tissue (M4) for 24 h. The nuclear maturation of DOs was assessed by the extrusion of the first polar body and the cytoplasmic maturation was evaluated by subsequently developmental capacity after parthenogenetic activation. More DOs matured to MII (56.89%) and developed to blastocysts (25.75%) when they were matured in vitro with M3 in comparison with DOs matured in vitro with M1 (45.14 and 15.97%) and M4 (40.48 and 13.49%). Further detection of gap junctions by injecting Lucifer yellow directly into cytoplasm of matured DOs with adherent cumulus cells and scanning with confocal microscope showed that Lucifer yellow were found in nine out of 11 the adherent cumulus cells in M3, indicating that the gap junctions between oocytes and cumulus cells was reconstructed in vitro. These results indicate that co-culture of DOs embedded in cumulus cell clumps can improve their nuclear and cytoplasmic maturation of DOs, possibly through the reconstruction of gap junctions in vitro.

The earliest stages of folliculogenesis in vitro

Reproduction ◽  
2002 ◽  
pp. 185-202 ◽  
Author(s):  
JE Smitz ◽  
RG Cortvrindt
Keyword(s):  
Ovarian Tissue ◽  
Ovarian Follicle ◽  
Mammalian Species ◽  
Follicle Development ◽  
Follicle Growth ◽  
Follicle Culture ◽  
In Vitro Techniques ◽  
Suitable Material ◽  
Culture Systems

In recent years several follicle culture systems have been pioneered in different mammalian species for studying ovarian folliculogenesis and culturing immature oocytes. Applications of these in vitro techniques include fertility preservation for humans, conservation of rare animals and development of oocyte banks for research purposes. Immature female gametes in the ovarian cortex can be cryopreserved for later use if culture techniques are available afterwards to promote growth and maturation. This review focuses on biochemical and biophysical factors related to oocyte culture in mice, the only animal in which live offspring have been produced after folliculogenesis in vitro. The advantage of using mice for these studies is that, in parallel to development of follicle culture systems, essential knowledge on folliculogenesis can be obtained from knockout mouse models. Recent experiments in mice stressed the principal role of the oocyte in follicle development and the strict timing of the biological processes underlying oogenesis in vitro. In large domestic animals and humans, study of oocyte culture is confounded by the constitutively prolonged nature of ovarian follicle development. In humans, only some aspects of follicle development have been studied because of the limited availability of suitable material for experimentation, technical difficulties related to manipulation of very small structures and lack of knowledge on physiological regulation of the early stages of follicle growth. Only a few reports describe ovarian follicular growth in vitro. In this review, relevant information on hormonal and growth factor regulation of the earliest stages of follicle growth in mammals is reviewed. Techniques are becoming available for the precise isolation of distinct classes of follicle and powerful molecular biology techniques can be used in studies of ovarian tissue culture.

scholarly journals Curcumin in Combination with Aerobic Exercise Improves Follicular Dysfunction via Inhibition of the Hyperandrogen-Induced IRE1α/XBP1 Endoplasmic Reticulum Stress Pathway in PCOS-Like Rats

2021 ◽  
Vol 2021 ◽  
pp. 1-22
Author(s):  
Yaling Zhang ◽  
Yajing Weng ◽  
Daojuan Wang ◽  
Rong Wang ◽  
Lihui Wang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Aerobic Exercise ◽  
Ovarian Function ◽  
Polycystic Ovary ◽  
Ovarian Tissue ◽  
Oral Intake ◽  
Follicle Development ◽  
Sprague Dawley

Combining diet with exercise can improve health and performance. Exercise can reduce androgen excess and insulin resistance (IR) in polycystic ovary syndrome (PCOS) patients. Curcumin is also presumed to improve the follicle development disorder. Here, we investigated the effects of a combination therapy of oral intake of curcumin and exercise on hyperandrogen-induced endoplasmic reticulum (ER) stress and ovarian granulosa cell (GC) apoptosis in rats with PCOS. We generated a PCOS model via continuous dehydroepiandrosterone subcutaneous injection into the necks of Sprague Dawley rats for 35 days. PCOS-like rats then received curcumin treatment combined with aerobic (treadmill) exercise for 8 weeks. We found that compared to control rats, the ovarian tissue and ovarian GCs of hyperandrogen-induced PCOS rats showed increased levels of ER stress-related genes and proteins. Hyperandrogen-induced ovarian GC apoptosis, which was mediated by excessive ER stress and unfolded protein response (UPR) activation, could cause follicle development disorders. Both curcumin gavage and aerobic exercise improved ovarian function via inhibiting the hyperandrogen-activated ER stress IRE1α-XBP1 pathway. Dihydrotestosterone- (DHT-) induced ER stress was mitigated by curcumin/irisin or 4μ8C (an ER stress inhibitor) in primary GC culture. In this in vitro model, the strongly expressed follicular development-related genes Ar, Cyp11α1, and Cyp19α1 were also downregulated.

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