In vitro development of pig preantral follicles cultured in a serum-free medium and the effect of angiotensin II

Shuttleworth, Gail; Pipkin, Fiona Broughton; Hunter, M. G.

doi:10.1530/rep.0.1230807

Cited by 43 publications

(11 citation statements)

References 21 publications

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“…Follicles containing enzymatically isolated primordial follicles, freely cultured or grown in collagen gels, are burdened by collapse, migration of cumulus granulosa away from the follicle, or oocyte degeneration [22,25,26]. Enzymatic digestion also disrupts the theca cell layer and basement membrane [27], despite efforts to limit exposure to enzymes [28]. The development of methods to isolate intact primordial follicles will allow utilization of the significant primordial follicular pool for human IVM and fertility preservation.…”

Section: Discussionmentioning

confidence: 99%

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

Krotz

Robins

Ferruccio

et al. 2010

J Assist Reprod Genet

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Purpose Create a 3-Dimensional artificial human ovary to mature human oocytes. Methods Theca and granulosa cells were isolated from antral follicles of reproductive-aged women, seeded into micro-molded gels and self-assembled into complex 3D microtissues. Immunohistochemistry and live-dead staining confirmed theca cell identity and cellular viability at one week respectively. Placement of granulosa cell spheroids or cumulus-oocyte complexes into theca cell honeycomb openings resulted in creation of an artificial human ovary. Oocytes from this construct were assessed for polar body extrusion.Results Theca and granulosa cells self-assembled into complex microtissues, remaining viable for one week. At 72 h after artificial human ovary construction, theca cells completely surrounded the granulosa spheroids or COCs without stromal invasion or disruption. Polar body extrusion occurred in one of three COCs assessed. Conclusions An artifical human ovary can be created with self-assembled human theca and granulosa cell microtissues, and used for IVM and future oocyte toxicology studies.

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Section: Discussionmentioning

confidence: 99%

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

Krotz

Robins

Ferruccio

et al. 2010

J Assist Reprod Genet

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“…However, the Ang II type 1 and 2 receptors (AGTR1 and AGTR2) have been identified in porcine granulosa cells of earlier stages of follicle development (Shuttleworth et al 2002b). A positive action of Ang II in estradiol synthesis and growth of preantral follicles in culture were also reported in swine (Shuttleworth et al 2002a). The hypothesis that RAS is involved in early follicle development was recently reinforced.…”

Section: Ang II Receptors In Follicular Cellsmentioning

confidence: 99%

Role of angiotensin in ovarian follicular development and ovulation in mammals: a review of recent advances

Gonçalves¹,

Ferreira²,

Gasperin³

et al. 2012

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Angiotensin (Ang) II is widely known for its role in the control of systemic blood vessels. Moreover, Ang II acts on the vascular control of ovarian function, corpus luteum formation, and luteolysis. Over the past 10 years, our research group has been studying the new concept of the renin-angiotensin system (RAS) as an autocrine/paracrine factor regulating steroidogenesis and promoting different cellular responses in the ovary, beyond vascular function. We have developed and used different in vivo and in vitro experimental models to study the role of RAS in the ovary and a brief overview of our findings is presented here. It is widely accepted that there are marked species differences in RAS function in follicle development. Examples of species-specific functions of the RAS in the ovary include the involvement of Ang II in the regulation of follicle atresia in rats vs the requirement of this peptide for the dominant follicle development and ovulation in rabbits and cattle. More recently, Ang-(1-7), its receptor, and enzymes for its synthesis (ACE2, NEP, and PEP) were identified in bovine follicles, implying that Ang-(1-7) has an ovarian function. Other novel RAS components (e.g. (pro)renin receptor and renin-binding protein) recently identified in the bovine ovary show that ovarian RAS is poorly understood and more complex than previously thought. In the present review, we have highlighted the progress toward understanding the paracrine and autocrine control of ovarian antral follicle development and ovulation by ovarian tissue RAS, focusing on in vivo studies using cattle as a model.

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“…Preantral follicles were mechanically isolated, with the aid of a stereomicroscope, as described previously (Cecconi et al 1999, Shuttleworth et al 2002. After isolation, healthy follicles were selected on the basis of their morphology (Shuttleworth et al 2002).…”

Section: Oocytes Collectionmentioning

confidence: 99%

“…After isolation, healthy follicles were selected on the basis of their morphology (Shuttleworth et al 2002). Follicle diameters were measured with the aid of an inverted phase microscope equipped with an ocular micrometer (40!…”

Section: Oocytes Collectionmentioning

confidence: 99%

Vascular supply as a discriminating factor for pig preantral follicle selection

Martelli¹,

Bernabò²,

Berardinelli³

et al. 2009

Reproduction

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This research analyses how somatic and vascular compartments change during preantral follicle growth. To address this aim, theca-granulosa (somatic) proliferation indexes (PIs), proportion of proliferating endothelial cells (PE), vascular area (VA) and vascular endothelial growth factor A (VEGFA) expression were simultaneously recorded on single healthy preantral follicles, classified into six different stages on the basis of the diameter and the granulosa layers. An autonomous blood vessel network starts to appear only in class 3. Vascular remodelling requires VEGFA expression, and VEGFA mRNA and VA significantly increase between class 3 and classes 4 and 5 and, further, in class 6. In addition, a positive correlation exists between these parameters in classes 3-5. Despite variation in angiogenesis results from classes 3 to 5, the statistical analysis reveals that the vascular parameters are positively and strictly correlated with somatic PIs. Conversely, class 6, also characterized by higher values of somatic PIs, displays a stable proportion of PEs (y40%) without showing any correlation among the different parameters analysed. To identify follicular subpopulations within different classes, a multivariate hierarchical cluster analysis was performed. This analysis reveals that the majority of classes 3 and 4 are quiescent follicles or structures that grow very slowly. Class 5 represents a transitory category, where half of the follicles maintain a low activity and the remaining express significantly higher levels of granulosa PI and VA. The follicles with this high activity are probably able to reach class 6 becoming dominant structures where somatic and vascular parameters are constantly on high levels and the VA remains the unique differentiating element.

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In vitro development of pig preantral follicles cultured in a serum-free medium and the effect of angiotensin II

Cited by 43 publications

References 21 publications

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

Role of angiotensin in ovarian follicular development and ovulation in mammals: a review of recent advances

Vascular supply as a discriminating factor for pig preantral follicle selection

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