Vitrification preserves murine ovarian follicular cell transcriptome in a 3D encapsulated in vitro follicle growth system

2021 ◽  
Author(s):  
Yingzheng Wang ◽  
Daniela D Russo ◽  
Pawat Pattarawat ◽  
Qiang Zhang ◽  
Mary B Zelinski ◽  
...  
Keyword(s):  
Ovarian Follicle ◽  
Principal Component ◽  
Follicular Cell ◽  
Molecular Signatures ◽  
Follicle Growth ◽  
Expression Of Genes ◽  
Robust Model ◽  
Cell Transcriptome ◽  
Conservation Of Endangered Species

Abstract Vitrification is a method for long-term biological sample cryopreservation without causing intra- and extra-cellular ice formation. We recently established a novel closed vitrification system to cryopreserve mouse ovarian follicles. Using the 3D alginate hydrogel encapsulated in vitro follicle growth (eIVFG) method, we demonstrated that compared to freshly-harvested follicles, vitrified follicles had normal follicle and oocyte reproductive outcomes. However, it is unknown whether vitrification preserves molecular signatures of folliculogenesis, which is the primary research focus in this study. Six fresh and six vitrified antral follicles grown from eIVFG were collected on day 8 for the whole single-follicle RNA sequencing. Principal component analysis (PCA) and Pearson’s correlation analysis revealed that vitrified follicles had similar transcriptomic profiles to fresh follicles. There were 35 differentially expressed genes between vitrified and fresh follicles, however, none of those genes have been shown to be critical to folliculogenesis and oogenesis. Meanwhile, gene ontology (GO) and KEGG pathway analysis revealed that no GO terms or signaling pathways were significantly enriched. Furthermore, the expression of genes essential for the gonadotropin-dependent folliculogenesis and oogenesis were comparable between vitrified and fresh follicles. Taken together, these results demonstrate that vitrification preserves follicular cell transcriptome and molecular signatures of gonadotropin-dependent folliculogenesis in the eIVFG system, providing a robust model for fertility preservation, conservation of endangered species, and also establishing a high-content ovarian follicle biobank for studying ovarian biology and female reproductive toxicology.

scholarly journals Involvement of hyaluronan synthesis in ovarian follicle growth in rats

Reproduction ◽  
2014 ◽  
Vol 147 (2) ◽  
pp. 189-197 ◽  
Author(s):  
Noriyuki Takahashi ◽  
Wataru Tarumi ◽  
Bunpei Ishizuka
Keyword(s):  
Ovarian Follicle ◽  
Primary Site ◽  
Corpora Lutea ◽  
Follicular Atresia ◽  
Follicle Growth ◽  
Adult Rats ◽  
Antral Follicles ◽  
Cell Layers ◽  
Ha Synthase

Most of the previous studies on ovarian hyaluronan (HA) have focused on mature antral follicles or corpora lutea, but scarcely on small preantral follicles. Moreover, the origin of follicular HA is unknown. To clarify the localization of HA and its synthases in small growing follicles, involvement of HA in follicle growth, and gonadotropin regulation of HA synthase (Has) gene expression, in this study, perinatal, immature, and adult ovaries of Wistar-Imamichi rats were examined histologically and biochemically and byin vitrofollicle culture. HA was detected in the extracellular matrix of granulosa and theca cell layers of primary follicles and more advanced follicles. Ovarian HA accumulation ontogenetically started in the sex cords of perinatal rats, and its primary site shifted to the intrafollicular region of primary follicles within 5 days of birth. TheHas1–3mRNAs were expressed in the ovaries of perinatal, prepubertal, and adult rats, and the expression levels ofHas1andHas2genes were modulated during the estrous cycle in adult rats and following administration of exogenous gonadotropins in immature acyclic rats. TheHas1andHas2mRNAs were predominantly localized in the theca and granulosa cell layers of growing follicles respectively. Treatments with chemicals known to reduce ovarian HA synthesis induced follicular atresia. More directly, the addition ofStreptomyceshyaluronidase, which specifically degrades HA, induced the arrest of follicle growth in anin vitroculture system. These results indicate that gonadotropin-regulated HA synthesis is involved in normal follicle growth.

Influence of gonadotropins on ovarian follicle growth and developmentin vivoandin vitro

Zygote ◽  
2017 ◽  
Vol 25 (3) ◽  
pp. 235-243 ◽  
Author(s):  
Maxim Filatov ◽  
Yulia Khramova ◽  
Elena Parshina ◽  
Tatiana Bagaeva ◽  
Maria Semenova
Keyword(s):  
Luteinizing Hormone ◽  
Assisted Reproductive Technology ◽  
Ovarian Follicle ◽  
Follicle Stimulating Hormone ◽  
Reproductive Technology ◽  
Present Review ◽  
Follicle Growth ◽  
Therapeutic Practice ◽  
Gonadotropic Hormones

SummaryGonadotropins are the key regulators of ovarian follicles development. They are applied in therapeutic practice in assisted reproductive technology clinics. In the present review we discuss the basic gonadotropic hormones – recombinant human follicle-stimulating hormone, its derivatives, luteinizing hormone and gonadotropin serum of pregnant mares, their origin, and application in ovarian follicle systems inin vitroculture systems.

N-acetyl-cysteine and the control of oxidative stress during in vitro ovarian follicle growth, oocyte maturation, embryo development and cryopreservation

2021 ◽  
pp. 106801
Author(s):  
Laryssa G. Barrozo ◽  
Laís R.F.M. Paulino ◽  
Bianca R. Silva ◽  
Efigênia C. Barbalho ◽  
Danisvânia R. Nascimento ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Ovarian Follicle ◽  
Follicle Growth ◽  
Acetyl Cysteine

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.

Differential expression of signal transduction factors in ovarian follicle development: a functional role for betaglycan and FIBP in granulosa cells in cattle

2008 ◽  
Vol 33 (2) ◽  
pp. 193-204 ◽  
Author(s):  
N. Forde ◽  
M. Mihm ◽  
M. J. Canty ◽  
A. E. Zielak ◽  
P. J. Baker ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Growth Trajectories ◽  
Dominant Follicle ◽  
Quantitative Real Time Pcr ◽  
Differential Growth ◽  
Ovarian Follicle Development ◽  
Expression Of Genes

Ovarian follicles develop in groups yet individual follicles follow different growth trajectories. This growth and development are regulated by endocrine and locally produced growth factors that use a myriad of receptors and signal transduction pathways to exert their effects on theca and granulosa cells. We hypothesize that differential growth may be due to differences in hormonal responsiveness that is partially mediated by differences in expression of genes involved in signal transduction. We used the bovine dominant follicle model, microarrays, quantitative real-time PCR and RNA interference to examine this. We identified 83 genes coding for signal transduction molecules and validated a subset of them associated with different stages of the follicle wave. We suggest important roles for CAM kinase-1 and EphA4 in theca cells and BCAR1 in granulosa cells for the development of dominant follicles and for betaglycan and FIBP in granulosa cells of regressing subordinate follicles. Inhibition of genes for betaglycan and FIBP in granulosa cells in vitro suggests that they inhibit estradiol production in regressing subordinate follicles.

Corrigendum to: Hypoxia limits mouse follicle growth in vitro

2017 ◽  
Vol 29 (2) ◽  
pp. 431 ◽  
Author(s):  
J. M. Connolly ◽  
M. T. Kane ◽  
L. R. Quinlan ◽  
P. Dockery ◽  
A. C. Hynes
Keyword(s):  
Ascorbic Acid ◽  
Ovarian Follicle ◽  
Time Frame ◽  
Maximum Diameter ◽  
Mouse Serum ◽  
Follicle Growth ◽  
Follicle Culture ◽  
Serum Type

Ovarian follicle culture is useful for elucidation of factors involved in the regulation of follicular function. We examined the effects of gas phase oxygen concentration, an oil overlay, serum type and medium supplementation with FSH, insulin–transferrin–selenium (ITS) and I-ascorbic acid on cultured preantral mouse follicle growth in a spherical, non-attached follicle culture system. Follicle growth in 5% oxygen was significantly (PPP>0.05) affected by an oil overlay, ITS supplementation or serum type. Culture in medium with 5% mouse serum, 1 IU mL–1 FSH, 25 μgmL–1 l-ascorbic acid and 20% oxygen without an oil overlay supported the growth of follicles to a maximum diameter of 380 μm in 6 days. Compared with mature preovulatory mouse follicles in vivo that often have diameters >500 μm within the same time frame, in vitro-grown follicles clearly exhibit limited growth. Thus, adequate oxygenation is an essential factor in the process of optimising follicle growth.

scholarly journals Melatonin as an endogenous hormone to slow down the exhaustion of ovarian follicle reserve in mice–a novel insight into its roles in early folliculogenesis and ovarian aging

2020 ◽  
Author(s):  
Chan Yang ◽  
Qinghua Liu ◽  
Yingjun Chen ◽  
Xiaodong Wang ◽  
Zaohong Ran ◽  
...  
Keyword(s):  
Ovarian Follicle ◽  
Primordial Follicle ◽  
High Dose ◽  
Follicle Growth ◽  
Ovarian Aging ◽  
Neonatal Mice ◽  
Primordial Follicle Activation ◽  
Follicle Activation

Abstract Previous studies have shown that long-term intake of exogenous melatonin can effectively delay ovarian aging, but the mechanism has not been fully elucidated. We observed that SNAT, the rate-limiting enzyme in the melatonin synthetic pathway, is localized in primordial and early follicle, and that granulosa cells isolated from follicle can synthesize melatonin. In vitro cultured neonatal mice ovaries with melatonin inhibited primordial follicle activation and early follicle growth. In vivo experiments further indicated that daily injections of melatonin to neonatal mice during the primordial follicle activation phase can reduce the number of activated follicles by inhibiting the PI3K-AKT-FOXO3 pathway; during the early follicle growth phase, injections of melatonin significantly suppressed early follicle growth and atresia, and transcriptome data showed that multiple pathways involved in folliculogenesis, including PI3K-AKT, were suppressed. Further, SNAT knockout in mice resulted in a significant increase in follicle activation and atresia, and eventually accelerated ovarian aging. We also demonstrated that prolonged high-dose melatonin intake had no obvious adverse effect on the health condition of mice. This study confirms that endogenous melatonin is involved in the regulation of ovarian aging, and reveals that melatonin delays ovarian aging by inhibiting primordial follicle activation, early follicle growth and atresia.

Effect of heifer age on the granulosa cell transcriptome after ovarian stimulation

2018 ◽  
Vol 30 (7) ◽  
pp. 980 ◽  
Author(s):  
David A. Landry ◽  
Rémi Labrecque ◽  
François-Xavier Grand ◽  
Christian Vigneault ◽  
Patrick Blondin ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Bos Taurus ◽  
Gene Expression Pattern ◽  
Principal Component ◽  
Developmental Competence ◽  
Improvement Programs ◽  
Breed Improvement ◽  
Cell Transcriptome ◽  
Sexually Mature

Genomic selection is accelerating genetic gain in dairy cattle. Decreasing generation time by using younger gamete donors would further accelerate breed improvement programs. Although ovarian stimulation of peripubertal animals is possible and embryos produced in vitro from the resulting oocytes are viable, developmental competence is lower than when sexually mature cows are used. The aim of the present study was to shed light on how oocyte developmental competence is acquired as a heifer ages. Ten peripubertal Bos taurus Holstein heifers underwent ovarian stimulation cycles at the ages of 8, 11 (mean 10.8) and 14 (mean 13.7) months. Collected oocytes were fertilised in vitro with spermatozoa from the same adult male. Each heifer served as its own control. The transcriptomes of granulosa cells recovered with the oocytes were analysed using microarrays. Differential expression of certain genes was measured using polymerase chain reaction. Principal component analysis of microarray data revealed that the younger the animal, the more distinctive the gene expression pattern. Using ingenuity pathway analysis (IPA) and NetworkAnalyst (www.networkanalyst.ca), the main biological functions affected in younger donors were identified. The results suggest that cell differentiation, inflammation and apoptosis signalling are less apparent in peripubertal donors. Such physiological traits have been associated with a lower basal concentration of LH.

Sign in / Sign up

Export Citation Format

Share Document