Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 5/2010

01-05-2010 | Fertility Preservation

In vitro culture of sheep lamb ovarian cortical tissue in a sequential culture medium

Authors: Xiayu Peng, Mei Yang, Liqin Wang, Chen Tong, Zhiqin Guo

Published in: Journal of Assisted Reproduction and Genetics | Issue 5/2010

Login to get access

Abstract

Purpose

This study evaluates the effect of a sequential culture system on the follicular development of sheep lamb ovaries, aiming to establish an available in vitro culture system for ovarian culture.

Methods

Lamb ovarian cortical fragments were cultured on a steel mesh with a nitrocellulose membrane pre-coated by type1 collagen. Several culture media were used for the determinations, specifically, a control medium (α-MEM), a constant medium (control medium supplemented with 75 ng/mL human recombinant EGF, 200 mIU/mL sheep FSH, 100 ng/mL human recombinant GDF-9, and 100 ng/mL human recombinant bFGF), and a sequential medium (control medium supplemented with sequential growth factors added on different days). Ovarian tissues, both fresh and cultured, were processed for histological and apoptotic assays, while spent culture media were processed for hormone assays.

Results

It was found that the growth of lamb primordial follicles can be initiated during culture in vitro. Compared to the control medium, sequential culture medium significantly increased the percentage of secondary follicles in cultures, while the follicle and oocyte diameters of primary and secondary follicles were also observed to increase in this medium. The constant medium was found to increase the number and diameter of secondary follicles only 18 days after culture. After this same period of time, some normal antral follicles were found in the sequential medium, while a few abnormal antral-like follicles were found in the control medium. Moreover, sequential medium appeared to significantly increase estradiol and inhibin production, especially 10–18 days after culture. The highest percentage of normal follicles and the lowest apoptotic cell rates were observed in the sequential medium, suggesting that a sequential addition style of culture can improve follicle and tissue viability.

Conclusions

The sequential addition of FSH, EGF, GDF-9, and bFGF can stimulate primordial follicle transmittal into the later development stages, even as far as the antral stage, improve the survival rate of follicles, and maintain follicular viability.
Literature
1.
Nayudu PL, Vitt UA, de Tomasi Jorgelina Barrios, Katti Pancharatna, Alfredo Ulloa-Aguirre. Intact follicle culture: what it can tell us about the roles of FSH glycoforms during follicle development. Reprod BioMed Online. 2002;5:240–53.CrossRefPubMed
2.
Adriaens I, Cortvrindt R, Smitz J. Differential FSH exposure in preantral follicle culture has marked effects on folliculogenesis and oocyte developmental competence. Hum Reprod. 2004;19:398–408.CrossRefPubMed
3.
Matos MHT, Lima-Verde IB, Luque MCA, Maia Jr JE, Silva JRV, Celestino JJH, et al. Essential role of follicle stimulating hormone in the maintenance of caprine preantral follicle viability in vitro. Zygote. 2007;15:173–82.CrossRefPubMed
4.
Hayashi M, McGee EA, Min G, Klein C, Rose UM, van Duin Marcel, et al. Recombinant growth differentiation factor-9 (GDF-9) enhances growth and differentiation of cultured early ovarian follicles. Endocrinology. 1999;140:1236–44.CrossRefPubMed
5.
Vitt UA, Hayashi M, Klein C, Hsueh AJ. Growth differentiation factor-9 stimulates proliferation but suppresses the follicle-stimulating hormone-induced differentiation of cultured granulosa cells from small antral and preovulatory rat follicles. Biol Reprod. 2000;62:370–7.CrossRefPubMed
6.
Hreinsson JG, Scott JE, Rasmussen C, Swahn ML, Hsueh AJ, Hovatta O. Growth differentiation factor-9 promotes the growth, development, and survival of human ovarian follicles in organ culture. [J] Clin Endocrinol Metab. 2002;87:316–21.CrossRef
7.
Martins FS, Celestino JJH, Saraiva MVA, Matos MHT, Bruno JB, Rocha-Junior MC, et al. Growth and differentiation factor-9 stimulates activation of goat primordial follicles in vitro and their progression to secondary follicles. Reprod Fertil Dev. 2008;20:916–24.CrossRefPubMed
8.
Nilsson EE, Skinner MK. Kit ligand and basic fibroblast growth factor interactions in the induction of ovarian primordial to primary follicle transition. Mol Cell Endocrinol. 2004;214:19–25.CrossRefPubMed
9.
Matos MH, Lima-Verde IB, Bruno JB, Lopes CA, Martins FS, Santos KD, et al. Follicle stimulating hormone and fibroblast growth factor-2 interact and promote goat primordial follicle development in vitro. Reprod Fertil Dev. 2007;19:677–84.CrossRefPubMed
10.
Roni G, Ronit A, Arye E, Carmela F, Shmuel N, Benjamin F. Effects of basic fibroblast growth factor on in vitro development of human ovarian primordial follicles. Fertil Steril. 2009;91:1967–75.CrossRef
11.
Kezele PR, Nilsson EE, Skinner MK. Insulin but not insulin-like growth factor-1 promotes the primordial to primary follicle transition. Mol Cell Endocrinol. 2002;192:37–43.CrossRefPubMed
12.
Boland NI, Gosden RG. Effects of epidermal growth factor on the growth and differentiation of cultured mouse ovarian follicles. J Reprod Fertil. 1994;101:369–74.PubMed
13.
Wandji SA, Eppig JJ, Fortune JE. FSH and growth factors affect the growth and endocrine function in vitro of granulosa cells of bovine preantral follicles. Theriogenology. 1996;45:817–32.CrossRefPubMed
14.
Gutierrez CG, Ralph JH, Telfer EE, Wilmut I, Webb R. Growth and antrum formation of bovine preantral follicles in long-term culture in vitro. Biol Reprod. 2000;62:1322–8.CrossRefPubMed
15.
Telfer EE, McLaughlin M, Ding C, Thong KJ. A two-step serum-free culture system supports development of human oocytes from primordial follicles in the presence of activin. Hum Reprod. 2008;23(5):1151–8.CrossRefPubMed
16.
Cushman RA, Wahl CM, Fortune JE. Bovine ovarian cortical pieces grafted to chick embryonic membranes: a model for studies on the activation of primordial follicles. Hum Reprod. 2002;17:48–54.CrossRefPubMed
17.
Hovatta O, Silye R, Abir R, Krausz T, Winston RM. Extracellular matrix improves survival of both stored and fresh human primordial and primary ovarian follicles in long-term culture. Hum Reprod. 1997;12:1032–6.CrossRefPubMed
18.
Scott JE, Carlsson IB, Bavister BD, Hovatta O. Human ovarian tissue cultures: extracellular matrix composition, coating density and tissue dimensions. Reprod BioMed. 2004;9(3):287–93.CrossRef
19.
Oktay K, Newton H, Mullan J, Gosden RG. Development of human primordial follicles to antral stages in SCID/hpg mice stimulated with follicle stimulating hormone. Hum Reprod. 1998;13:1133–8.CrossRefPubMed
20.
Silva JR, van den Hurk R, Costa SH, Andrade ER, Nunes AP, Ferreira FV, et al. Survival and growth of goat primordial follicles after in vitro culture of ovarian cortical slices in media containing coconut water. Anim Reprod Sci. 2004;81:273–86.CrossRefPubMed
21.
Kezele P, Skinner MK. Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone: endocrine model of follicle assembly. Endocrinology. 2003;144:3329–37.CrossRefPubMed
22.
Durlinger AL, Gruijters MJ, Kramer P, Karels B, Ingraham HA, Nachtigal MW, et al. Anti-Mullerian hormone inhibits initiation of primordial follicle growth in the mouse ovary. Endocrinology. 2002;143:1076–84.CrossRefPubMed
23.
Derrar N, Price CA, Sirard MA. Effect of growth factors and co-culture with ovarian medulla on the activation of primordial follicles in explants of bovine ovarian cortex. Theriogenology. 2000;54:587–98.CrossRefPubMed
24.
Braw-Tal R, Yossefi S. Studies in vivo and in vitro on the initiation of follicle growth in the bovine ovary. J Reprod Fertil. 1997;109:165–71.PubMedCrossRef
25.
Fortune JE, Kito S, Wandji AS, Srsen V. Activation of bovine and baboon primordial follicles in vitro. Theriogenology. 1998;49:441–9.CrossRefPubMed
26.
Wright CS, Hovatta O, Margara R, Trew G, Winston RML, Franks S, et al. Effects of follicle-stimulating hormone and serum substitution on the in vitro growth of human ovarian follicles. Hum Reprod. 1999;14:1555–62.CrossRefPubMed
27.
Wang J, Roy SK. Growth differentiation factor-9 and stem cell factor promote primordial follicle formation in the hamster: modulation by follicle-stimulating hormone. Biol Reprod. 2004;70:577–85.CrossRefPubMed
28.
Nilsson E, Parrott JA, Skinner MK. Basic fibroblast growth factor induces primordial follicle development and initiates folliculogenesis. Mol Cell Endocrinol. 2001;175:123–30.CrossRefPubMed
29.
Silva JRV, van den Hurk R, de Matos MHT, dos Santos RR, Pessoa C, de Moraes MO, et al. Influences of FSH and EGF on primordial follicles during in vitro culture of caprine ovarian cortical tissue. Theriogenology. 2004;61:1691–704.CrossRefPubMed
30.
Huanmin Zhou, Yong Zhang. Regulation of in vitro growth of preantral follicles by growth factors in goats. Domest Anim Endocrinol. 2005;28:235–42.CrossRef
31.
Markstrom E, Svensson E, Shao R, Svanberg B, Billig H. Survival factors regulating ovarian apoptosis-dependence on follicle differentiation. Reproduction. 2002;123:23–30.CrossRefPubMed
32.
Cortvrindt R, Smitz J, Van Steirteghem AC. Assessment of the need for follicle stimulating hormone in early preantral mouse follicle culture in vitro. Hum Reprod. 1997;12:759–68.CrossRefPubMed
33.
Van den Hurk R, Abir R, Telfer EE, Bevers MM. Primate and bovine immature oocytes and follicles as source of fertilizable oocytes. Hum Reprod. 2000;6:457–74.CrossRef
34.
Shikone T, Yamoto M, Nakano R. Follicle-stimulating hormone induces functional receptors for basic fibroblast growth factor in rat granulosa cells. Endocrinology. 1992;131:1063–8.CrossRefPubMed
35.
Hreinsson JG, Scott JE, Rasmussen C, Swaha ML, Hsueh AJW, Hovatta O. Growth Differentiation Factor-9 Promotes the Growth, Development, and Survival of Human Ovarian Follicles in Organ Culture. J Clin Endocrinol Metab. 2002;87(1):316–21.CrossRefPubMed
36.
Rohan RM, Rexroad J, Guthrie HD. Changes in the concentration of MRNAS for the inhibin subunits in ovarian follicles after administration of gonadotropins to progestin-treated ewes. Domest Anim Endocrinol. 1991;8:445–54.CrossRefPubMed
37.
Souza CJH, Campbell BK, McNeilly AS, Baird DT. Effect of bone morphogenetic protein 2 (BMP2) on oestradiol and inhibin A production by sheep granulosa cells, and localization of BMP receptors in the ovary by immunohistochemistry. Reproduction. 2002;123:363–9.CrossRefPubMed
38.
Fortune JE. The early stages of follicular development: activation of primordial follicles and growth of preantral follicles. Anim Reprod Sci. 2003;78:135–63.CrossRefPubMed
39.
Behl R, Pandey RS. Effect of epidermal growth factor on steroidogenesis by caprine granulosa cells in culture: interaction with FSH. Small Rumin Res. 2001;40:57–62.CrossRefPubMed
40.
Wang Haibin, Xia Guoliang, Li Meiling, Guo Yong, Xie Huirong. Effects of Epidermal Growth Factor on the Follicle Development and Estradiol Secretion of Mouse Fetal Ovary Induced by Follicle-Stimulating Hormone in vitro. Journal of Agricultural Biotechnology. 2001;9:37–40.
41.
Elvin JA, Clark AT, Wang P, Wolfman NM, Matzuk MM. Paracrine actions of growth differentiation factor-9 in the mammalian ovary. Mol Endocrinol. 1999;13:1035–48.CrossRefPubMed
42.
Roh JS, Bondestam J, Mazerbourg S, Kaivo-Oja N, Groome N, Ritvos O, et al. Growth differentiation factor-9 stimulates inhibin production and activates Smad2 in cultured rat granulosa cells. Endocrinology. 2003;144(1):172–8.CrossRefPubMed
43.
Vernon RK, Spicer LJ. Effects of basic fibroblast growth factor and heparin on follicle-stimulating hormone-induced steroidogenesis by bovine granulosa cells. J Anim Sci. 1994;72:2696–702.PubMed
44.
Torrance C, Telfer E, Gosden RG. Quantitative study of the development of isolated mouse pre-antral follicles in collagen gel culture. J Reprod Fertil. 1989;87:367–74.PubMedCrossRef
45.
Pangas SA, Saudye H, Shea LD, Woodruff TK. Novel approach for the three-dimensional culture of granulosa cell-oocyte complexes. Tissue eng. 2003;9:1013–21.CrossRefPubMed
Metadata
Title
In vitro culture of sheep lamb ovarian cortical tissue in a sequential culture medium
Authors
Xiayu Peng
Mei Yang
Liqin Wang
Chen Tong
Zhiqin Guo
Publication date
01-05-2010
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 5/2010
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-010-9415-6

Other articles of this Issue 5/2010

Journal of Assisted Reproduction and Genetics 5/2010 Go to the issue

Abstracts

Meeting abstracts

Proceedings Summary

Proceedings of the First World Congress on Fertility Preservation: executive summary

Fertility Preservation

Using decision trees to enhance interdisciplinary team work: the case of oncofertility

Erratum

Erratum to: Oocyte cryopreservation: is it time to remove its experimental label?

Fertility Preservation

In vitro maturation, apoptotic gene expression and incidence of numerical chromosomal abnormalities following cryotop vitrification of sheep cumulus-oocyte complexes

Commentary

On meetings that matter