Culture of human preovulatory granulosa cells: Effect of extracellular matrix on steroidogenesis

Bussenot, Isabelle; Ferre, Guy; Azoulay-Barjonet, Catherine; Murgo, Christine; Vieitez, G.; Parinaud, Jean

doi:10.1016/s0248-4900(05)80186-8

Cited by 16 publications

(14 citation statements)

References 24 publications

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“…In our model, we can suggest that the drastic cell rounding triggered by addition of the anti-6 IgG directly affected GC steroidogenesis. Accordingly, previous studies have also reported an enhancement of E2 secretion by bovine or human GC exhibiting a rounded rather than a flattened shape (Ben-Rafael et al 1988, Bussenot et al 1993, Gutiérrez et al 1997. However, in rat (Carnegie & Tsang 1988), human (Ben-Rafael et al 1988) and pig (Sites et al 1996), in contrast to sheep, a rounded cell shape has also been associated with an enhancement of P4 secretion by GC.…”

Section: Discussionmentioning

confidence: 92%

Laminin-alpha6beta1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells

Bellego¹,

Pisselet²,

Huet³

et al. 2002

Journal of Endocrinology

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This study aimed to determine the physiological role of laminin (LN) and its receptor, 6 1 integrin, in controlling the functions of granulosa cells (GC) during follicular development in sheep ovary. Immunohistochemistry experiments showed the presence of increasing levels of LN (P<0·0001), and high levels of mature 6 1 integrin in GC layers of healthy antral follicles during the follicular and the preovulatory phases of the estrous cycle. In vitro, the addition of a function-blocking antibody raised against 6 subunit (anti-6 IgG) to the medium of ovine GC cultured on LN impaired cell spreading (P<0·0001), decreased the proliferation rate (P<0·05) and increased the apoptosis rate (P<0·05). Furthermore, addition of anti-6 IgG enhanced estradiol (E2) secretion by GC in the presence or absence of follicle-stimulating hormone (FSH), luteinizing hormone or insulin-like growth factor-I in culture medium (P<0·0001), and inhibited progesterone (P4) secretion in basal conditions or in the presence of low (0·5 ng/ml) FSH concentrations only (P<0·0001). The anti-6 IgG effect was specific to an interaction of LN with 6 1 integrin since it was ineffective on GC cultured on heat-denatured LN, RGD (arginine-glycine-aspartic acid) peptides and non-coated substratum. Hence, this study established that 6 1 integrin 1) was expressed in GC of antral follicles, 2) mediated the actions of LN on survival, proliferation and steroidogenesis of GC, and 3) was able to dramatically modulate P4 and E2 secretion by GC in vitro. It is suggested that during the follicular and the preovulatory phases of the estrous cycle, the increasing levels of LN in GC of large antral follicles might support their final development to ovulation.

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Section: Discussionmentioning

confidence: 92%

Laminin-alpha6beta1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells

Bellego¹,

Pisselet²,

Huet³

et al. 2002

Journal of Endocrinology

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“…Previous studies have shown that type I collagen stimulates GC steroidogenesis in vitro, in terms of the secretion of estradiol (human, Bussenot et al 1993), progesterone (rat, cow, Luck et al 1991), or both steroids (human, Ben-Rafael et al 1988). The discrepancies in the natures of the steroids induced by type I collagen may be due to differences between species or between the initial states of differentiation of GCs recovered for culture.…”

Section: Discussionmentioning

confidence: 98%

“…Similarly, luteinized human GCs exhibiting a rounded shape when cultured on a collagen matrix show increased estradiol secretion (Ben-Rafael et al 1988, Bussenot et al 1993. Notably, the rounded shape has been also associated with an enhancement of progesterone secretion in rat , human (Ben-Rafael et al 1988) and pig (Sites et al 1996) GCs cultured on ECM, suggesting that the relationship between cell shape and steroidogenesis is not unequivocal.…”

Section: Introductionmentioning

confidence: 99%

Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis: relationships between cell shape and function

Huet¹,

Pisselet²,

Mandon‐Pepin³

et al. 2001

Journal of Endocrinology

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The extracellular matrix (ECM), constituting the follicular basal lamina and present also between follicular cells and in the follicular fluid, is believed to regulate granulosa cell (GC) function during follicular development. Ovine GCs isolated from small (1-3 mm in diameter) or large (4-7 mm in diameter) antral follicles were cultured on various pure ECM components (type I collagen, fibronectin, laminin), synthetic substrata enhancing (RGD peptides) or impairing (poly 2-hydroxyethylmethacrylate (poly-hema)) cell adhesion, or in the presence of heparin. The effects of these factors, used alone or in combination with IGF-I and/or FSH, were evaluated in terms of GC spread, survival, proliferation and steroidogenesis.When grown on type I collagen (CI) gel, poly-hema or heparin, GCs from both large and small follicles exhibited a round shape and a low proliferation rate. Compared with non-coated plastic substratum as a control, these ECM or synthetic compounds enhanced estradiol secretion and reduced progesterone secretion by large-follicle GCs. In contrast, GCs from both large and small follicles spread extensively on CI coating, fibronectin, laminin and RGD peptides. Fibronectin and laminin dramatically increased the proliferation rate and enhanced survival of GCs from both origins. Moreover, fibronectin, laminin and RGD peptides reduced estradiol secretion by large-follicle GCs. Unexpectedly, CI coating increased estradiol secretion and reduced progesterone secretion by large-follicle GCs, suggesting that type I collagen was able to maintain estradiol secretion independently of GC shape. Finally, GC responsiveness to IGF-I and FSH, in terms of proliferation and steroidogenesis, was generally maintained when cells were grown on ECM components, RGD peptides and in the presence of heparin. However, when large-follicle GCs were grown as non-adherent clusters (as observed on poly-hema) basal and IGF-I-and/or FSH-stimulated progesterone secretions were totally abolished.Overall, this study shows that GC shape, survival, proliferation and steroidogenesis can be modulated in vitro by pure ECM components in a specific and coordinated manner. It is suggested that, in vivo, fibronectin and laminin would sustain follicular development by enhancing the survival and proliferation of GCs, whereas type I collagen might participate in the maintenance of estradiol secretion in large antral follicles.

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“…Reports with other animal models have not indicated type I collagen localization to the OSE, yet freshly isolated rat OSE and an immortal OSE line was shown to produce Type I collagen (Auersperg et al 1991). Type I collagen has been widely used with in vitro cultures of granulosa cells and follicles (Carnegie et al 1988;Bussenot et al 1993;Huet et al 2001). Type I collagen gels affect the shape and steroidogenesis of granulosa cells.…”

Section: Discussionmentioning

confidence: 99%

Distribution of extracellular matrix proteins type I collagen, type IV collagen, fibronectin, and laminin in mouse folliculogenesis

Berkholtz

Lai

Woodruff

et al. 2006

Histochem Cell Biol

132

113

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The extracellular matrix (ECM) plays a prominent role in ovarian function by participating in processes such as cell migration, proliferation, growth, and development. Although some of these signaling processes have been characterized in the mouse, the relative quantity and distribution of ECM proteins within developing follicles of the ovary have not been characterized. This study uses immunohistochemistry and real-time PCR to characterize the ECM components type I collagen, type IV collagen, fibronectin, and laminin in the mouse ovary according to follicle stage and cellular compartment. Collagen I was present throughout the ovary, with higher concentrations in the ovarian surface epithelium and follicular compartments. Collagen IV was abundant in the theca cell compartment with low-level expression in the stroma and granulosa cells. The distribution of collagen was consistent throughout follicle maturation. Fibronectin staining in the stroma and theca cell compartment increased throughout follicle development, while staining in the granulosa cell compartment decreased. Heavy staining was also observed in the follicular fluid of antral follicles. Laminin was localized primarily to the theca cell compartment, with a defined ring at the exterior of the follicular granulosa cells marking the basement membrane. Low levels of laminin were also apparent in the stroma and granulosa cell compartment. Taken together, the ECM content of the mouse ovary changes during follicular development and reveals a distinct spatial and temporal pattern. This understanding of ECM composition and distribution can be used in the basic studies of ECM function during follicle development, and could aid in the development of in vitro systems for follicle growth.

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Culture of human preovulatory granulosa cells: Effect of extracellular matrix on steroidogenesis

Cited by 16 publications

References 24 publications

Laminin-alpha6beta1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells

Laminin-alpha6beta1 integrin interaction enhances survival and proliferation and modulates steroidogenesis of ovine granulosa cells

Extracellular matrix regulates ovine granulosa cell survival, proliferation and steroidogenesis: relationships between cell shape and function

Distribution of extracellular matrix proteins type I collagen, type IV collagen, fibronectin, and laminin in mouse folliculogenesis

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