Differential regulation of pig theca cell steroidogenesis by LH, insulin-like growth factor I and granulosa cells in serum-free culture

Shores, EM; Picton, HM; Hunter, M. G.

doi:10.1530/reprod/118.2.211

Cited by 16 publications

(13 citation statements)

References 2 publications

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“…It has been previously used to assess the viability of porcine oocytes, granulosa and theca cells [50,51]. In the present study, the viability of follicular cells in tissue strips was assessed, respectively, at day 0 and at day 7 of in vitro culture in both fresh and vitrified-warmed groups.…”

Section: Evaluation Of Follicles In Situ With Neutral Redmentioning

confidence: 99%

In vitro development of human primordial follicles to preantral stage after vitrification

Khosravi

Reid

Moini

et al. 2013

J Assist Reprod Genet

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Purpose The aim was to culture primordial follicles in vitro to reach preantral stage in vitrified human ovarian tissue. Methods Ovarian tissue samples were obtained from six women. Tissue strips were vitrified by infiltration with a cryoprotectant followed by mounting on a stainless steel carrier. After culturing for 7 days the morphology and developmental stages of follicles enclosed in fresh and vitrified groups were analyzed. Results High proportion of viable follicles in vitrified ovarian strips was obtained. After culturing for 7 days the percentage of secondary and preantral follicles increased significantly (P <0.05) whereas primordial and transitory follicles showed a significant decrease (P <0.05) compared to their respective counterparts at day 0 of culture.Conclusions Vitrification of ovarian strips with an improved carrier device and culturing of follicles in ovarian strips after warming yielded developed follicles with high viability and morphological integrity that may be suitable for use in fertility preservation among cancer patients.

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Section: Evaluation Of Follicles In Situ With Neutral Redmentioning

confidence: 99%

In vitro development of human primordial follicles to preantral stage after vitrification

Khosravi

Reid

Moini

et al. 2013

J Assist Reprod Genet

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“…The theca pieces were cut into small pieces and enzymatically dispersed using Hank's balanced salt solution supplemented with 20 mM Hepes, 0.5% w/v collagenase type II, 0.1% w/v hyaluronidase type I-S and 5% fetal calf serum in a shaking water bath at 37°C [30]. The digested cell suspension was washed by centrifugation and resuspended in culture medium: DMEM/Ham's F12 with Hepes and sodium bicarbonate supplemented with 3 mM L-glutamine, 4 ng/ml selenium, 2.5 μg/ml transferrin, 0.1% BSA, 100 IU/ml penicillin and 0.1 mg/ml streptomycin.…”

Section: Methodsmentioning

confidence: 99%

“…50 × 10 3 viable cells in 50 μl aliquots were seeded into each well of a 96-well plate which contained 200 μl of culture medium supplemented with 10 ng/ml bovine insulin, 100 ng/ml Long R3 IGF-1 and 0.01 ng/ml porcine LH (USDA-pLH-B-2). An optimal dose of LR3 IGF-1 (100 ng/ml) used in granulosa cell cultures was also found to be optimal for theca cell function [30], hence the utilization of control cultures with 100 ng/ml LR3 IGF-1. Shores et al [30] observed that a combination of LR3 IGF-1 (100 ng/ml), insulin (10 ng/ml) and LH (0.01 ng/ml) were required to maintain viable thecal cell numbers in vitro .…”

Section: Methodsmentioning

confidence: 99%

“…An optimal dose of LR3 IGF-1 (100 ng/ml) used in granulosa cell cultures was also found to be optimal for theca cell function [30], hence the utilization of control cultures with 100 ng/ml LR3 IGF-1. Shores et al [30] observed that a combination of LR3 IGF-1 (100 ng/ml), insulin (10 ng/ml) and LH (0.01 ng/ml) were required to maintain viable thecal cell numbers in vitro .…”

Section: Methodsmentioning

confidence: 99%

“…Culture vessels have been coated with attachment factors [26], which can also result in luteinization; and different populations of granulosa cells (cumulus versus mural cells) have been incubated. Primary serum-free culture systems have been developed for ovine [27], bovine [28] and porcine [29] granulosa cells and also for porcine theca cells [30]. The advantage of these serum free culture systems is that they maintain the follicular phenotype in vitro , in that spontaneous luteinization is avoided and the granulosa cells are able to maintain the activity of the aromatase enzyme complex [27-29].…”

Section: Introductionmentioning

confidence: 99%

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Paracrine effects of oocyte secreted factors and stem cell factor on porcine granulosa and theca cells in vitro

Brankin

Mitchell

Webb

et al. 2003

Reprod Biol Endocrinol

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Oocyte control of granulosa and theca cell function may be mediated by several growth factors via a local feedback loop(s) between these cell types. This study examined both the role of oocyte-secreted factors on granulosa and thecal cells, cultured independently and in co-culture, and the effect of stem cell factor (SCF); a granulosa cell derived peptide that appears to have multiple roles in follicle development. Granulosa and theca cells were isolated from 2–6 mm healthy follicles of mature porcine ovaries and cultured under serum-free conditions, supplemented with: 100 ng/ml LR3 IGF-1, 10 ng/ml insulin, 100 ng/ml testosterone, 0–10 ng/ml SCF, 1 ng/ml FSH (granulosa), 0.01 ng/ml LH (theca) or 1 ng/ml FSH and 0.01 ng/ml LH (co-culture) and with/without oocyte conditioned medium (OCM) or 5 oocytes. Cells were cultured in 96 well plates for 144 h, after which viable cell numbers were determined. Medium was replaced every 48 h and spent medium analysed for steroids.Oocyte secreted factors were shown to stimulate both granulosa cell proliferation (P < 0.001) and oestradiol production (P < 0.001) by granulosa cells throughout culture. In contrast, oocyte secreted factors suppressed granulosa cell progesterone production after both 48 and 144 hours (P < 0.001). Thecal cell numbers were increased by oocyte secreted factors (P = 0.02), together with a suppression in progesterone and androstenedione synthesis after 48 hours (P < 0.001) and after 144 hours (P = 0.02), respectively. Oocyte secreted factors also increased viable cell numbers (P < 0.001) in co-cultures together with suppression of progesterone (P < 0.001) and oestradiol (P < 0.001). In granulosa cell only cultures, SCF increased progesterone production in a dose dependent manner (P < 0.001), whereas progesterone synthesis by theca cells was reduced in a dose dependent manner (P = 0.002). Co-cultured cells demonstrated an increase in progesterone production with increasing SCF dose (P < 0.001) and an increase in oestradiol synthesis at the highest dose of SCF (100 ng/ml). In summary, these findings demonstrate the presence of a co-ordinated paracrine interaction between somatic cells and germ cells, whereby oocyte derived signals interact locally to mediate granulosa and theca cell function. SCF has a role in modulating this local interaction. In conclusion, the oocyte is an effective modulator of granulosa-theca interactions, one role being the inhibition of luteinization.

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Role of granulosa and theca cell interactions in ovarian follicular maturation

Tajima

Orisaka

Yata

et al. 2006

Microscopy Res & Technique

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We developed a culture system in which two types of ovarian follicular cells were allowed to attach to opposite sides of a collagen membrane. Using this in vitro cell culture system, we studied the effects of granulosa- and theca-cell interaction on the morphology, structure, and function of bovine ovarian follicular cells. In the first part of the study, we explored how the interaction between theca and granulosa cells affects the morphology and structure of the cells. This study was done using follicular cells collected from bovine ovarian follicles at the early developmental stage. Granulosa cells cultured alone were flattened, and formed a monolayer sheet. By contrast, granulosa cells cultured with theca cells were convex, and formed multilayer sheets. Theca cells cultured alone were thin, flat, and spindle-shaped. Theca cells cultured with granulosa cells were also spindle-shaped; however, they appeared convex and more densely packed when compared with theca cells cultured alone. In the second part of the study, the possible role of the cellular interaction in the control of differentiation and growth of granulosa and theca cells was investigated. When follicular cells were isolated from the early stage of follicular development, theca cells reduced progesterone and inhibin production by granulosa cells and augmented the growth of granulosa cells. When the cells were isolated from the late stage of follicular development, by contrast, theca cells augmented hormonal production by granulosa cells, and did not affect the growth of granulosa cells. The growth and androstenedione production by theca cells were increased by the presence of granulosa cells, irrespective of the origin of follicular cells. These results demonstrated that communication between two types of follicular cells results in reciprocal modulation of their morphology, structure, growth, and function. Cellular interactions seem to be one of the major factors controlling the differentiation and growth of the follicular cells during the follicular maturation process. In contrast to granulosa and theca cells cultured alone, cells in the coculture seemed to possess morphological and functional characteristics more similar to those of cells in the growing follicular wall in vivo. Thus, we speculate that the interaction between these two types of follicular cells is essential for the maintenance of original structure and function of the bovine follicular wall.

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Differential regulation of pig theca cell steroidogenesis by LH, insulin-like growth factor I and granulosa cells in serum-free culture

Cited by 16 publications

References 2 publications

In vitro development of human primordial follicles to preantral stage after vitrification

In vitro development of human primordial follicles to preantral stage after vitrification

Paracrine effects of oocyte secreted factors and stem cell factor on porcine granulosa and theca cells in vitro

Role of granulosa and theca cell interactions in ovarian follicular maturation

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