Growth and histology of ovarian follicles after cold storage in the tammar wallaby

2006 ◽  
Vol 18 (6) ◽  
pp. 677 ◽  
Author(s):  
Nadine M. Richings ◽  
Geoffrey Shaw ◽  
Peter D. Temple-Smith ◽  
Marilyn B. Renfree
Keyword(s):  
Cold Storage ◽  
Ovarian Tissue ◽  
Tammar Wallaby ◽  
Ovarian Follicles ◽  
Interstitial Tissue ◽  
Similar Amount ◽  
Obvious Effect ◽  
Simple Method ◽  
Histological Techniques

Cold storage is a simple method for storing and transporting tissues and organs. The reliability of this method for maintaining structure and function of marsupial ovarian tissue was assessed using histological techniques and follicle culture. Tammar wallaby ovaries were placed in cold storage (phosphate-buffered saline at 4°C) for 24 or 48 h. Although necrotic changes were evident in the germinal epithelium, cortex and interstitial tissue after cold storage, there was little evidence of necrotic changes in ovarian follicles and oocytes appeared normal. Secondary follicles isolated from ovarian tissue after cold storage grew by a similar amount to non-stored follicles when cultured for 4 days in vitro, but no follicles from any group developed to tertiary follicles. Cold storage for up to 24 h had little obvious effect on the structure of ovarian tissue and follicles isolated from this tissue maintained their structure during culture. However, degeneration in culture increased with storage time and was significantly higher after cold storage for 48 h. As demonstrated in the tammar wallaby, cold storage has potential as a method for storage and transport of marsupial ovaries up to 24 h.

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.

Ovarian steroid metabolism and oestrogens in the corpus luteum of the tammar wallaby

1984 ◽  
Vol 101 (2) ◽  
pp. 231-NP ◽  
Author(s):  
M. B. Renfree ◽  
A. P. F. Flint ◽  
S. W. Green ◽  
R. B. Heap
Keyword(s):  
Corpus Luteum ◽  
Follicular Development ◽  
Hydroxysteroid Dehydrogenase ◽  
Tammar Wallaby ◽  
Steroid Metabolism ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Embryonic Diapause ◽  
Probable Source

ABSTRACT Ovaries were obtained from tammar wallabies at various stages of the reproductive cycle to examine the occurrence of oestrogens in corpora lutea, and the synthesis and metabolism of steroids in the corpus luteum and ovarian cortical and interstitial tissues. Corpora lutea contained oestradiol-17β and oestrone during embryonic diapause and at all stages of pregnancy studied after blastocyst activation. Aryl sulphatase, 3β-hydroxysteroid dehydrogenase and 17β-oxidoreductase were shown to be present in luteal and other ovarian tissues by incubation in vitro with labelled substrates. Aromatase was undetectable in corpora lutea or in interstitial tissue, but was present in the ovarian tissues (including follicles) which remained after removal of corpora lutea. The probable source of the oestrogens detected in the corpus luteum is discussed in relation to their role in the inhibition of follicular development during embryonic diapause. J. Endocr. (1984) 101, 231–240

Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Zygote ◽  
2015 ◽  
Vol 23 (6) ◽  
pp. 943-950 ◽  
Author(s):  
R.J.S. Gonçalves ◽  
A.Y.P. Cavalcante ◽  
B.B. Gouveia ◽  
T.L.B. Lins ◽  
R.S. Barberino ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Ovarian Follicles ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Oocyte Diameter ◽  
Minimal Essential Medium ◽  
Preantral Follicles ◽  
Apoptosis Rate ◽  
Physiological Values

SummaryThe aim of this study was to investigate the effect of ovarian tissue transportation conditions (medium and period of time) on the morphology, apoptosis and development of ovine preantral follicles cultured in vitro. Each ovarian pair was cut into nine slices, with one fragment being fixed immediately (fresh control). The remaining fragments were placed individually in cryotubes containing conservation medium (minimal essential medium (MEM) without supplementation or MEM+ – with supplementation) and stored at 35ºC for 6 or 12 h without (non-cultured) or with subsequent culture for 5 days. Then, the fragments were processed for histological and terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labelling (TUNEL) examination. Preservation of ovarian slices in MEM or MEM+ (non-cultured) resulted in similar percentages of normal follicles when compared with the fresh control. Nevertheless, compared with the fresh control, a decrease in the percentage of normal follicles was observed in tissues cultured for 5 days. Only for tissues preserved in supplemented medium (MEM+) for 6 h, the percentage of TUNEL positive cells was similar between non-cultured tissues and tissues cultured for 5 days. Follicular activation and growth (follicular and oocyte diameter) were higher in cultured tissues than in fresh control or non-cultured tissues, except those from fragments preserved for 6 h in MEM and then cultured for 5 days in which no growth was observed. In conclusion, ovine ovarian tissue was successfully preserved in supplemented medium (MEM+) at a temperature close to physiological values (35°C) for up to 6 h without affecting apoptosis in the ovarian follicles and their ability to develop in vitro.

scholarly journals METABOLIC AND ULTRASTRUCTURAL CHANGES IN THE FROG OVARIAN FOLLICLE IN RESPONSE TO PITUITARY STIMULATION

1970 ◽  
Vol 46 (3) ◽  
pp. 491-504 ◽  
Author(s):  
John Walberg Anderson ◽  
Milton B. Yatvin
Keyword(s):  
Epithelial Cells ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Ovarian Tissue ◽  
Ovarian Follicle ◽  
Ovarian Follicles ◽  
Ultrastructural Changes ◽  
Surface Epithelium

Frog ovarian fragments were prevented from ovulating in vitro by the addition of actinomycin D up to 3 hr following pituitary stimulation; but addition of Actinomycin D 6 hr after stimulation was far less effective. Puromycin, on the other hand, effectively inhibited ovulation when added as late as 6 hr after pituitary stimulation. Although actinomycin D reduced uptake of uridine-3H, and puromycin reduced uptake of leucine-3H and lysine-14 by pituitary-stimulated ovarian tissue minus oocytes (OTMO) in vitro, it was found that pituitary stimulation did not significantly increase uptake of these compounds by OTMO. Radioautographs of ovarian follicles fixed 6 hr after the addition of pituitary extract and uridine-3H in vitro revealed increased RNA synthesis in the peritoneal surface epithelium, compared with unstimulated controls, while the ovarian sac epithelium showed no increase. Gross ultrastructural changes occurred in the peritoneal area of ovarian follicles following pituitary stimulation in vivo, including loss of collagen fibrils, and general disorganization of the connective tissue theca. Changes in the rough endoplasmic reticulum of the peritoneal epithelial cells, while frequently encountered, were less pronounced. None of these changes was observed in the ovarian sac area, or in the interfollicular region. The above data are consistent with the hypothesis that pituitary stimulation of the frog ovary results in increased synthesis of RNA and protein by the peritoneal epithelial cells, and that the protein may be collagenase.

Incorporation of arginine, glutamine or leucine in culture medium accelerates in vitro activation of primordial follicles in 1-day-old mouse ovary

Zygote ◽  
2020 ◽  
Vol 28 (5) ◽  
pp. 409-416
Author(s):  
Parimah Alborzi ◽  
Mohammad Jafari Atrabi ◽  
Vahid Akbarinejad ◽  
Ramezan Khanbabaei ◽  
Rouhollah Fathi
Keyword(s):  
Amino Acids ◽  
Culture Medium ◽  
Ovarian Tissue ◽  
Mammalian Target Of Rapamycin ◽  
Primordial Follicle ◽  
Ovarian Follicles ◽  
Primordial Follicles ◽  
Base Medium ◽  
Elevated Expression

SummaryIn vitro activation of primordial follicles provides cancer patients subjected to oncotherapy with a safe therapeutic strategy for fertility preservation, however a successful protocol for activation of primordial follicles in prepubertal patients has not yet been defined comprehensively. There is evidence that amino acids such as leucine, arginine and glutamine could stimulate the mammalian target of rapamycin (mTOR) pathway, which plays a pivotal role in primordial follicle activation. Nevertheless, there has been no report that elucidates the effect of these amino acids on in vitro development of ovarian follicles. Therefore, the present study was conducted to evaluate the effects of these amino acids and their combination on the formation and activation of primordial follicles in 1-day-old murine ovaries during an 11-day culture period. The experimental groups consisted of base medium (BM), base medium + arginine (ARG), base medium + glutamine (GLU), base medium + leucine (LEU) and base medium + a combination of arginine, glutamine and leucine (AGL). The proportions of different stages of ovarian follicles and gene expression of regulatory factors were assessed using histology and quantitative real-time PCR on days 5 and 11 of culture. The proportion of transitional and primary follicles was greater in all amino acid-treated groups compared with the BM group (P < 0.05). Moreover, leucine resulted in elevated expression of Gdf9 and Bmp15, and glutamine augmented the expression of Pi3k on day 11 of culture. In conclusion, the present study showed that inclusion of leucine, glutamine, arginine or their combination in the culture medium for murine ovarian tissue could accelerate the activation of primordial follicles and alter the expression of the corresponding factors.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

A new in vitro model for pre-transplantation diagnosis of frozen/thawed human ovarian tissue

2011 ◽  
Vol 71 (05) ◽  
Author(s):  
M Salama ◽  
K Winkler ◽  
KF Murach ◽  
S Hofer ◽  
L Wildt ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Ovarian Tissue ◽  
Human Ovarian Tissue ◽  
Vitro Model

The Filter Loop Technique as a Method of Measuring Platelet Aggregation in the Flowing Blood of the Rat; the Inhibitory Activity of 5-oxo-l-cyclopentene-l-heptanoic Acid (AY-16,804) on Platelet Aggregation

1973 ◽  
Vol 30 (01) ◽  
pp. 138-147 ◽  
Author(s):  
Christopher R. Muirhead
Keyword(s):  
Platelet Aggregation ◽  
Prostaglandin E1 ◽  
Suitable Model ◽  
Heptanoic Acid ◽  
Simple Method ◽  
Loop Technique ◽  
Flowing Blood ◽  
Filter Loop

SummaryThe filter loop technique which measures platelet aggregation in vivo in the flowing-blood of the rat was compared to the optical density technique of Born which is carried out in vitro with platelet rich plasma. Using these two experimental models the effect on platelet aggregation of three known inhibitors sulfinpyrazone, dipyridamole and prostaglandin E1, and a novel compound 5-oxo-l-cyclopentene-l-heptanoic acid (AY-16, 804) was determined.The effects on platelet aggregation of the known inhibitors were consistent with information in the literature. Prostaglandin E1 was the most potent inhibitor in both techniques; sulfinpyrazone inhibited aggregation in both models but was less potent than prostaglandin E1. AY-16, 804 exhibited activity in vitro and in vivo similar to that of sulfinpyrazone. Dipyridamole did not inhibit platelet aggregation in vivo and did not inhibit aggregation in vitro in concentrations at which it remained soluble.The filter loop technique is a suitable model for measuring platelet aggregation in the flowing blood of the rat. It is a relatively simple method of determining aggregation and easily adapted to other species.

THE AUTORADIOGRAPHIC DISTRIBUTION PATTERN AFTER ADMINISTRATION OF DIETHYLSTILBOESTROL COMPARED WITH THAT OF NATURAL OESTROGENS

1963 ◽  
Vol 43 (4) ◽  
pp. 561-570 ◽  
Author(s):  
Gösta Bengtsson ◽  
Sven Ullberg
Keyword(s):  
Distribution Pattern ◽  
Adrenal Cortex ◽  
Natural Compounds ◽  
Ovarian Follicles ◽  
Interstitial Tissue ◽  
Specific Accumulation ◽  
Natural And Synthetic

ABSTRACT The distribution in mice of 14C- and 3H-diethylstilboestrol has been investigated autoradiographically. The results have been compared with those which have been previously reported for natural oestrogens. Many similarities have been demonstrated between the synthetic and natural compounds. Thus a specific accumulation has been observed in the endometrium, the granulosa layer of large ovarian follicles, the adrenal cortex. the interstitial tissue of the testes, and the hypophysis. Natural and synthetic oestrogens differ widely concerning the penetration into and the distribution within the foetus.

STEROID FORMATION IN PORCINE OVARIAN TISSUE IN VITRO

1970 ◽  
Vol 63 (3) ◽  
pp. 441-453 ◽  
Author(s):  
Asbjørn Aakvaag
Keyword(s):  
Liquid Scintillation ◽  
Specific Activity ◽  
Ovarian Tissue ◽  
Gas Liquid Chromatography ◽  
Similar Concentration ◽  
Endogenous Substrates ◽  
Exogenous Progesterone ◽  
Relative Utilization ◽  
Radioactive Substrate

ABSTRACT Slices of non-luteinized porcine ovaries have been incubated in the presence or absence of human chorionic gonadotrophin (HCG) and exogenous radioactive substrates. Progesterone, 17α-hydroxyprogesterone and androstenedione were isolated in a radiochemically pure form. The chemical mass and the specific activity were determined by gas liquid chromatography and liquid scintillation spectrometry. HCG stimulated the rate of formation of androstenedione in the absence of exogenous substrates with a factor of 4–8. In the presence of pregnenolone or progesterone at a concentration of about 2 × 10−6 mol/l the stimulatory effect of HCG was either abolished or markedly reduced. The conversion of exogenous progesterone to androstenedione was reduced in response to HCG indicating that the capacity of the tissue to convert progesterone to androstenedione was limited, and that the limit was reached at this rather low substrate concentration. These findings furthermore suggest that the endogenous rather than the exogenous radioactive substrate will be »preferred« by the tissue. The observations demonstrate the necessity of measuring both the radioactivity and the chemical mass of the products in investigations of this type using radioactive substrates. The formation of progesterone from endogenous substrates was also stimulated by HCG. [1-14C] acetate and [7α-3H]cholesterol were not utilized by the tissue for steroid formation. Exogenous [4-14C] pregnenolone and [7α-3H] progesterone in similar concentration were both utilized for production of 17α-hydroxyprogesterone and androstenedione. HCG had no effect on the relative utilization of the radioactive substrates.

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