Three-dimensional environments preserve extracellular matrix compartments of ovarian follicles and increase FSH-dependent growth

Co-culturing of follicles with interstitial cells in collagen gel reproduce follicular development accompanied with theca cell layer formation

“…Consistent with previous reports [19-25], the extracellular matrix proteins fibronectin (Figure 4c), laminin (Figure 4d), tenascin (Figure 4e), and collagen type IV (Figure 4f) were detected in the theca cell layers of the follicles in mouse ovaries. As reported previously [18], in addition to these extracellular matrix proteins, theca cell layers included the cell adhesion molecule THY1 (Figure 4g). In addition, the steroid synthesizing enzyme CYP17A-1, which is reported to be localized only in the theca cells in ovaries [3], was selectively detected in a subpopulation of the theca cells in mouse ovaries (Figure 4h).…”

“…We confirmed that the follicles maintained their three-dimensional shapes when cultured in collagen gel by measuring the vertical and horizontal diameters of serial sections (Figure 5), and using confocal microscopy [18]. Some follicle culture systems utilize collagen gel as the matrix.…”

Co-culturing of follicles with interstitial cells in collagen gel reproduce follicular development accompanied with theca cell layer formation

“…Consistent with previous reports [19-25], the extracellular matrix proteins fibronectin (Figure 4c), laminin (Figure 4d), tenascin (Figure 4e), and collagen type IV (Figure 4f) were detected in the theca cell layers of the follicles in mouse ovaries. As reported previously [18], in addition to these extracellular matrix proteins, theca cell layers included the cell adhesion molecule THY1 (Figure 4g). In addition, the steroid synthesizing enzyme CYP17A-1, which is reported to be localized only in the theca cells in ovaries [3], was selectively detected in a subpopulation of the theca cells in mouse ovaries (Figure 4h).…”

“…We confirmed that the follicles maintained their three-dimensional shapes when cultured in collagen gel by measuring the vertical and horizontal diameters of serial sections (Figure 5), and using confocal microscopy [18]. Some follicle culture systems utilize collagen gel as the matrix.…”

A Novel Three-Dimensional Culture System Allows Prolonged Culture of Functional Human Granulosa Cells and Mimics the Ovarian Environment

“…The presence of Call-Exner bodies in ovarian follicles has been used as an index of GC proliferation. 24,25 Using Ki-67, 16,26 we were able to demonstrate that cell proliferation occurs in the follicular cells cultured in 3D conditions. During prolonged culture, most proliferating cells progressively became organized in structures reminiscent of ovarian follicles with cells staining for FSHR and LHR and surrounded by Coll IV.…”

Quality of Oocytes Derived from Vitrified Ovarian Follicles Cultured in Two- and Three-Dimensional Culture System in the Presence and Absence of Kit Ligand

“…Moreover, the three-dimensional culture system can maintain follicular structure with an in vivo -like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48…”

“…Moreover, the threedimensional culture system can maintain follicular structure with an in vivo-like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48 We propose that high production of steroid hormone (E2, P4, and DHEA) by follicles grown in the three-dimensional culture system may be due to a high level of androgen provided by theca cells, and accelerated increased aromatase activity, and increased granulosa cell numbers.…”

Co-culturing of follicles with interstitial cells in collagen gel reproduce follicular development accompanied with theca cell layer formation

“…Consistent with previous reports [19-25], the extracellular matrix proteins fibronectin (Figure 4c), laminin (Figure 4d), tenascin (Figure 4e), and collagen type IV (Figure 4f) were detected in the theca cell layers of the follicles in mouse ovaries. As reported previously [18], in addition to these extracellular matrix proteins, theca cell layers included the cell adhesion molecule THY1 (Figure 4g). In addition, the steroid synthesizing enzyme CYP17A-1, which is reported to be localized only in the theca cells in ovaries [3], was selectively detected in a subpopulation of the theca cells in mouse ovaries (Figure 4h).…”

“…We confirmed that the follicles maintained their three-dimensional shapes when cultured in collagen gel by measuring the vertical and horizontal diameters of serial sections (Figure 5), and using confocal microscopy [18]. Some follicle culture systems utilize collagen gel as the matrix.…”

A Novel Three-Dimensional Culture System Allows Prolonged Culture of Functional Human Granulosa Cells and Mimics the Ovarian Environment

“…The presence of Call-Exner bodies in ovarian follicles has been used as an index of GC proliferation. 24,25 Using Ki-67, 16,26 we were able to demonstrate that cell proliferation occurs in the follicular cells cultured in 3D conditions. During prolonged culture, most proliferating cells progressively became organized in structures reminiscent of ovarian follicles with cells staining for FSHR and LHR and surrounded by Coll IV.…”

Quality of Oocytes Derived from Vitrified Ovarian Follicles Cultured in Two- and Three-Dimensional Culture System in the Presence and Absence of Kit Ligand

“…Moreover, the three-dimensional culture system can maintain follicular structure with an in vivo -like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48…”

“…Moreover, the threedimensional culture system can maintain follicular structure with an in vivo-like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48 We propose that high production of steroid hormone (E2, P4, and DHEA) by follicles grown in the three-dimensional culture system may be due to a high level of androgen provided by theca cells, and accelerated increased aromatase activity, and increased granulosa cell numbers.…”

Co-culturing of follicles with interstitial cells in collagen gel reproduce follicular development accompanied with theca cell layer formation

“…Consistent with previous reports [19-25], the extracellular matrix proteins fibronectin (Figure 4c), laminin (Figure 4d), tenascin (Figure 4e), and collagen type IV (Figure 4f) were detected in the theca cell layers of the follicles in mouse ovaries. As reported previously [18], in addition to these extracellular matrix proteins, theca cell layers included the cell adhesion molecule THY1 (Figure 4g). In addition, the steroid synthesizing enzyme CYP17A-1, which is reported to be localized only in the theca cells in ovaries [3], was selectively detected in a subpopulation of the theca cells in mouse ovaries (Figure 4h).…”

“…We confirmed that the follicles maintained their three-dimensional shapes when cultured in collagen gel by measuring the vertical and horizontal diameters of serial sections (Figure 5), and using confocal microscopy [18]. Some follicle culture systems utilize collagen gel as the matrix.…”

A Novel Three-Dimensional Culture System Allows Prolonged Culture of Functional Human Granulosa Cells and Mimics the Ovarian Environment

“…The presence of Call-Exner bodies in ovarian follicles has been used as an index of GC proliferation. 24,25 Using Ki-67, 16,26 we were able to demonstrate that cell proliferation occurs in the follicular cells cultured in 3D conditions. During prolonged culture, most proliferating cells progressively became organized in structures reminiscent of ovarian follicles with cells staining for FSHR and LHR and surrounded by Coll IV.…”

Quality of Oocytes Derived from Vitrified Ovarian Follicles Cultured in Two- and Three-Dimensional Culture System in the Presence and Absence of Kit Ligand

“…Moreover, the three-dimensional culture system can maintain follicular structure with an in vivo -like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48…”

“…Moreover, the threedimensional culture system can maintain follicular structure with an in vivo-like basal lamina architecture that minimizes the disruption of follicles. 46 It was demonstrated that three-dimensional culture of preantral follicles using an extracellular matrix provided a better milieu for in vitro growth and survival of follicles when compared with the conventional two-dimensional culture system. 47,48 We propose that high production of steroid hormone (E2, P4, and DHEA) by follicles grown in the three-dimensional culture system may be due to a high level of androgen provided by theca cells, and accelerated increased aromatase activity, and increased granulosa cell numbers.…”